Process for preparing homocystein analogs useful as intermediates for compounds containing a fused bicyclic ring

ABSTRACT

Homocysteine analogs of the formula ##STR1## wherein P 1  is a nitrogen protecting group and R 6  is alkyl, substituted alkyl or benzyl are prepared by esterifying N-protected L-methionine, oxidizing, treating the resulting sulfoxide with an acid anhydride, treating the resulting product with an alkali metal hydroxide followed by removal of formaldehyde and the treatment with an acid anhydride or acid halide. The L-homocysteine analogs are useful as intermediates in the preparation of compounds containing a fused bicyclic ring.

RELATED APPLICATION

This application is a division of Ser. No. 238,764 filed May 5, 1994 nowU.S. Pat. No. 5,508,272.

BACKGROUND OF THE INVENTION

Captopril, (S)-1-(3-mercapto-2-methyl-1-oxopropyl)-L-proline, having thestructural formula ##STR2## is an orally active angiotensin convertingenzyme inhibitor useful for treating hypertension and congestive heartfailure. See Ondetti et al. U.S. Pat. No. 4,105,776.

Enalapril,(S)-1-[N-[1-(ethoxycarbonyl)-3-phenylpropyl]-L-alanyl]-L-proline, havingthe structural formula ##STR3## is also an orally active angiotensinconverting enzyme inhibitor. Enalapril contains the L-alanyl-L-prolinedipeptide. A related compound, lisinopril, also possesses oralangiotensin converting enzyme inhibitor activity and contains theL-lysyl-L-proline dipeptide. See Harris et al. U.S. Pat. No. 4,374,829.

Fosinopril sodium,(4S)-4-cyclohexyl-1-[[(R)-[(S)-1-hydroxy-2-methylpropoxy](4-phenylbutyl)phosphinyl]acetyl]-L-proline propionate (ester), sodiumsalt having the structural formula ##STR4## is also an orally activeangiotensin converting enzyme inhibitor useful for treatinghypertension. See Petrillo U.S. Pat. No. 4,337,201.

Haslanger et al. in U.S. Pat. No. 4,749,688 disclose treatinghypertension by administering neutral metalloendopeptidase inhibitorsalone or in combination with atrial peptides or angiotensin convertingenzyme inhibitors.

Neustadt in U.S. Pat. No. 5,075,302 disclose that mercaptoacyl aminolactams of the formula ##STR5## wherein Y includes propylene andbutylene, R¹ is lower alkyl, aryl or heteroaryl, and R² is hydrogen,lower alkyl, lower alkoxy lower alkyl, aryl-lower alkyl orheteroaryl-lower alkyl are endopeptidase inhibitors. Neustadt discloseemploying such compounds alone or in combination with angiotensinconverting enzyme inhibitors to treat cardiovascular diseases such ashypertension, congestive heart failure, edema, and renal insufficiency.

Delaney et al. U.K. Patent 2,207,351 disclose that endopeptidaseinhibitors produce diuresis and natriuresis and are useful alone or incombination with angiotensin converting enzyme inhibitors for thereduction of blood pressure. Delaney et al. include various mercapto andacylmercapto amino acids and dipeptides among their endopeptidaseinhibiting compounds.

Flynn et al. in European Patent Application 481,522 disclose dualinhibitors of enkephalinase and angiotensin converting enzyme of theformulas ##STR6## wherein n is zero or one and Z is O, S, --NR₆ -- or##STR7## Additional tricyclic dual inhibitors are disclosed byWarshawsky et al. in European Patent Applications 534,363, 534,396 and534,492.

Karanewsky et al. in U.S. Pat. Nos. 4,432,971 and 4,432,972 disclosephosphonamidate angiotensin converting enzyme inhibitors of the formula##STR8## wherein X is a substituted imino or amino acid or ester.

Karanewsky in U.S. Pat. No. 4,460,579 discloses angiotensin convertingenzyme inhibitors including those of the formula ##STR9## and in U.S.Pat. No. 4,711,884 discloses angiotensin converting enzyme inhibitorsincluding those of the formula ##STR10## wherein X is a thiazine orthiazepine.

SUMMARY OF THE INVENTION

This invention is directed to novel compounds containing a fusedbicyclic ring which are useful as angiotensin converting enzymeinhibitors. Some of these compounds also possess neutral endopeptidaseinhibitory activity. This invention is also directed to pharmaceuticalcompositions containing such selective or dual action inhibitors and themethod of using such compositions. This invention is also directed tothe process for preparing such novel compounds, novel intermediates, andprocesses for preparing such intermediates.

The novel fused bicyclic inhibitors of this invention include thosecompounds of the formula ##STR11## and pharmaceutically acceptable saltsthereof wherein: A is ##STR12## X is O or S--(O)_(t) ; R₁ and R₁₂ areindependently selected from hydrogen, alkyl, alkenyl, cycloalkyl,substituted alkyl, substituted alkenyl, aryl, substituted aryl,heteroaryl, cycloalkyl-alkylene-, aryl-alkylene-, substitutedaryl-alkylene-, and heteroaryl-alkylene- or R₁ and R₁₂ taken togetherwith the carbon to which they are attached complete a cycloalkyl ring ora benzofused cycloalkyl ring;

R₂ is hydrogen, ##STR13## or R₁₁ --S--; R₃, R₅ and R₇ are independentlyselected from hydrogen, alkyl, substituted alkyl, aryl-(CH₂)_(p) --,substituted aryl-(CH₂)_(p) --, heteroaryl-(CH₂)_(p) --, ##STR14## R₄ isalkyl, cycloalkyl-(CH₂)_(p) --, substituted alkyl, aryl-(CH₂)_(p) --,substituted aryl-(CH₂)_(p) --, or heteroaryl-(CH₂)_(p) --;

R₆ is alkyl, substituted alkyl, cycloalkyl-(CH₂)_(p) --, aryl-(CH₂)_(p)--, substituted aryl-(CH₂)_(p) --, or heteroaryl-(CH₂)_(p) --;

R₈ is hydrogen, lower alkyl, cycloalkyl, or phenyl;

R₉ is hydrogen, lower alkyl, lower alkoxy, or phenyl;

R₁₀ is lower alkyl or aryl-(CH₂)_(p) --;

R₁₁ is hydrogen, alkyl, substituted alkyl, cycloalkyl-(CH₂)_(p) --,aryl-(CH₂)_(p) --, substituted aryl-(CH₂)_(p) --, heteroaryl-(CH₂)_(p)--, or --S--R₁₁ completes a symmetrical disulfide wherein R₁₁ is##STR15## m is zero or one; Y is CH₂, S--(O)_(t) or O provided that Y isS--(O)_(t) or O only when m is one;

n is one or two;

p is zero or an integer from 1 to 6;

q is zero or an integer from 1 to 3;

r is zero or one; and

t is zero, one, or two.

DETAILED DESCRIPTION OF THE INVENTION

The term "alkyl" refers to straight or branched chain radicals having upto seven carbon atoms. The term "lower alkyl" refers to straight orbranched radicals having up to four carbon atoms and is a preferredsubgrouping for the term alkyl.

The term "substituted alkyl" refers to such straight or branched chainradicals of 1 to 7 carbons wherein one or more, preferably one, two, orthree, hydrogens have been replaced by a hydroxy, amino, cyano, halo,trifluoromethyl, --NH(lower alkyl), --N(lower alkyl)₂, lower alkoxy,lower alkylthio, or carboxy.

The terms "lower alkoxy" and "lower alkylthio" refer to such lower alkylgroups as defined above attached to an oxygen or sulfur.

The term "cycloalkyl" refers to saturated rings of 3 to 7 carbon atomswith cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl being mostpreferred.

The term "alkenyl" refers to straight or branched chain radicals of 3 to7 carbon atoms having one or two double bonds. Preferred "alkenyl"groups are straight chain radicals of 3 to 5 carbons having one doublebond.

The term "substituted alkenyl" refers to such straight or branchedradicals of 3 to 7 carbons having one or two double bonds wherein ahydrogen has been replaced by a hydroxy, amino, halo, trifluoromethyl,cyano, --NH(lower alkyl), --N(lower alkyl)₂, lower alkoxy, loweralkylthio, or carboxy.

The term "alkylene" refers to straight or branched chain radicals havingup to seven carbon atoms, i.e. --CH₂ --, --(CH₂)₂ --, --(CH₂)₃ --,--(CH₂)₄ --, ##STR16## etc.

The term "aryl" refers to phenyl, 1-naphthyl, and 2-naphthyl. The term"substituted aryl" refers to phenyl, 1-naphthyl, and 2-naphthyl having asubstituent selected from lower alkyl, lower alkoxy, lower alkylthio,halo, hydroxy, trifluoromethyl, amino, --NH(lower alkyl), or --N(loweralkyl)₂, and di- and tri-substituted phenyl, 1-naphthyl, or 2-naphthylwherein said substituents are selected from methyl, methoxy, methylthio,halo, hydroxy, and amino.

The term "heteroaryl" refers to unsaturated rings of 5 or 6 atomscontaining one or two O and S atoms and/or one to four N atoms providedthat the total number of hetero atoms in the ring is 4 or less. Theheteroaryl ring is attached by way of an available carbon or nitrogenatom. Preferred heteroaryl groups include 2-, 3-, or 4-pyridyl,4-imidazolyl, 4-thiazolyl, 2- and 3-thienyl, and 2- and 3-furyl. Theterm heteroaryl also includes bicyclic rings wherein the five or sixmembered ring containing O, S, and N atoms as defined above is fused toa benzene or pyridyl ring. Preferred bicyclic rings are 2- and 3-indolyland 4- and 5-quinolinyl. The mono or bicyclic heteroaryl ring can alsobe additionally substituted at an available carbon atom by a loweralkyl, halo, hydroxy, benzyl, or cyclohexylmethyl. Also, if the mono orbicyclic ring has an available N-atom such N atom can also besubstituted by an N-protecting group such as ##STR17##2,4-dinitrophenyl, lower alkyl, benzyl, or benzhydryl.

The compounds of formula I wherein

A is ##STR18## X is O or S, and Y is CH₂, O, or S can be prepared bycoupling the acylmercapto containing sidechain of the formula ##STR19##with a fused bicyclic ring compound of the formula ##STR20## to give theproduct of the formula ##STR21## wherein R₃ is hydrogen or an acidprotecting group such as methyl, ethyl, t-butyl, or benzyl. The abovereaction can be performed in an organic solvent such as methylenechloride and in the presence of a coupling reagent such as1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicylcohexylcarbodiimide,benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate,or carbonyldiimidazole. Alternatively, the acylmercapto carboxylic acidof formula II can be converted to an activated form prior to couplingsuch as an acid chloride, mixed anhydride, symmetrical anhydride,activated ester, etc.

The product of formula IV can be converted to the mercaptan product offormula I wherein R₂ is hydrogen and R₃ is hydrogen by methods known inthe art. For example, when R₆ is methyl and R₃ is methyl or ethyltreatment with methanolic sodium hydroxide followed by aqueous acidyields the products wherein R₂ and R₃ are hydrogen.

The products of formula I wherein R₂ is hydrogen can be acylated with anacyl halide of the formula ##STR22## wherein halo is F, Cl or Br oracylated with an anhydride of the formula ##STR23## to give otherproducts of formula I wherein R₂ is ##STR24##

The products of formula I wherein R₂ is --S--R₁₁ and R₁₁ is alkyl,substituted alkyl, cycloalkyl-(CH₂)_(p) --, aryl-(CH₂)_(p) --,substituted aryl-(CH₂)_(p) --, or heteroaryl-(CH₂)_(p) -- can beprepared by reacting the products of formula I wherein R₂ is hydrogenwith a sulfonyl compound of the formula

    H.sub.3 C--SO.sub.2 --S--R.sub.11                          (VII)

in an aqueous alcohol solvent to yield the desired products. Thecompounds of formula VII are known in the literature or can be preparedby known methods, see for example, Smith et al., Biochemistry, 14, p766-771 (1975).

The product of formula I wherein R₂ is SH can be prepared by reactingthe product of formula I wherein R₂ is hydrogen with a compound offormula VII wherein R₁₁ is triphenylmethyl or trialkylsilyl followed byremoval of the triphenylmethyl or trialkylsilyl group under acidicconditions.

The symmetrical disulfide products of formula I can be prepared bydirect oxidation of the product of formula I wherein R₂ is hydrogen withiodine according to known procedures, see, for example, Ondetti et al.U.S. Pat. No. 4,105,776.

The acylmercapto sidechain compounds of formula II wherein R₁₂ ishydrogen are described in the literature. See, for example, Ondetti etal. U.S. Pat. Nos. 4,105,776 and 4,339,600, Haslanger et al. U.S. Pat.No. 4,801,609, Delaney et al. U.S. Pat. No. 4,722,810, etc.

The acylmercapto sidechain compounds of formula II wherein R₁ and R₁₂are both other than hydrogen and r is zero can be prepared by reactingthe substituted carboxylic acid of the formula ##STR25## withbis[[(4-methoxy)phenyl]methyldisulfide in the presence of lithiumdiisopropylamide to give the compound of the formula ##STR26## Treatmentof the compound of formula IX with strong acid such astrifluoromethanesulfonic acid removes the methoxybenzyl protecting groupand is followed by acylation with the acyl halide of formula V oranhydride of formula VI to give the compound of formula II wherein R₁and R₁₂ are both other than hydrogen and r is zero.

Alternatively, the substituted carboxylic acid of formula VIII can bereacted with lithium diisopropyl amide and sulfur to give the mercaptanof the formula ##STR27## The mercaptan of formula X can then be acylatedwith the acyl halide of formula V or the anhydride of formula VI to givethe compound of formula II wherein R₁ and R₁₂ are both other thanhydrogen and r is zero.

The acylmercapto sidechain compounds of formula II wherein R₁ and R₁₂are both other than hydrogen and r is one can be prepared by reactingthe substituted carboxylic acid of the formula ##STR28## withpara-toluenesulfonyl chloride in pyridine to give the lactam of theformula ##STR29## Treatment of the lactam of formula XII with a cesiumthioacid of the formula ##STR30## in the presence of dimethylformamideyields the desired acylmercapto sidechain of formula II wherein R₁ andR₁₂ are both other than hydrogen and r is one.

The compounds of formula I wherein A is ##STR31## X is O or S, and Y isCH₂, O, or S can be prepared by coupling the acid of the formula##STR32## wherein R₇ is an acid protecting group with the fused bicyclicring compound of formula III in the presence of a coupling reagent asdefined above to give the product of the formula ##STR33##Alternatively, the acid of formula XIV can be converted to an activatedform such as an acid chloride prior to the coupling reaction.

The acids of formula XIV are described by Warshawsky et al. in EuropeanPatent Application 534,396 and 534,492.

The compounds of formula I wherein A is ##STR34## X is O or S, and Y isCH₂, O, or S can be prepared by reacting a keto acid or ester of theformula ##STR35## with a fused bicyclic ring compound of formula IIIunder reducing conditions to give the product of the formula ##STR36##

The keto acids and esters of formula XVI are described in theliterature. See, for example, Ruyle U.S. Pat. No. 4,584,294 and Parsonset al. U.S. Pat. No. 4,873,235.

Alternatively, the fused bicyclic ring compound formula III can bereacted with a triflate of the formula ##STR37## to give the product offormula XVII.

The compounds of formula I wherein A is ##STR38## X is O or S, and Y isCH₂, O or S can be prepared by coupling a phosphonochloridate of theformula ##STR39## wherein R₅ is lower alkyl or benzyl with a fusedbicyclic ring compound of formula III to give the product of the formula##STR40## Preferably, R₃ in the compound of formula III is lower alkylor benzyl. The R₃ and R₅ acid protecting groups can then be removed, forexample, by hydrogenation to give the corresponding products of formulaI wherein R₃ and R₅ are hydrogen.

The phosphonochloridates of formula XIX are known in the literature.See, for example, Karanewsky et al. U.S. Pat. Nos. 4,432,971 and4,432,972 and Karanewsky U.S. Pat. No. 4,460,579.

The products of formula I wherein either x or Y or both are S--(O)_(t)and t is one or two can be prepared by oxidation of the compounds offormulas IV, XV, XVII, or XX with a known oxidizing reagent such as metachloroperbenzoic acid, peracetic acid, monoperoxyphthalic acid,magnesium salt hexahydrate, etc. By controlling the amount of oxidizingreagent and the time of the reaction, the products are obtained whereint is one or two.

The ester products of formula I wherein R₅ or R₇ is ##STR41## can beprepared by treating the corresponding compounds of formula I wherein R₅or R₇ is hydrogen and R₃ is an acid protecting group with a compound ofthe formula ##STR42## wherein L is a leaving group such as chloro,bromo, or tolylsulfonyloxy followed by removal of the R₃ esterprotecting group.

The ester products of formula I wherein R₃ is ##STR43## can be preparedby treating the corresponding compounds of formula I wherein R₃ ishydrogen and R₂ is ##STR44## with a compound of formula XXI.

The fused bicyclic ring compounds of formula III can be preparedaccording to the following processes which also form part of thisinvention. For example, when Y is CH₂ an N-protected amino acid of theformula ##STR45## can be coupled with the amino acid ester of theformula ##STR46## to give the dipeptide of the formula ##STR47## whereinP₁ is an amino protecting group such as benzyloxycarbonyl ort-butyloxycarbonyl or a group which together with the N-atom forms aprotecting group such as phthalimido, P₂ is a hydroxy or mercaptoprotecting group, and R₃ is an easily removable ester protecting group.Preferred P₂ protecting groups when X is S are acyl groups such asacetyl or benzoyl, especially acetyl. Preferred P₂ protecting groupswhen X is O are acyl groups, tetrahydropyrans, hindered silyl groups andtrityls, especially triphenylmethyl and 1,1-dimethylethyldimethylsilyl.This coupling reaction is preferably performed in the presence of acoupling reagent such asbenzotriazol-1-yloxytris-(dimethylamino)phosphonium hexafluorophosphate,ethyl-3-(3-dimethylamino)propyl carbodiimide, ormethanesulfonyloxybenzotriazole.

The P₂ protecting group can be selectively removed from the intermediateof formula XXIV such as by treatment with sodium methoxide in methanolwhen P₂ is acetyl or benzoyl or treatment with an acid such asp-toluenesulfonic acid in methanol when P₂ is acetyl, benzoyl, trityl,tetrahydropyranyl, or 1,1-dimethylethyldimethylsilyl. The resultingproduct is then subjected to an acid catalyzed cyclization reactionpreferably by treating with a strong acid such as trifluoroacetic acid,para-toluenesulfonic acid or a commercially available polystyrenesulfonate polymer type ion exchange resin such as Amberlyst 15®. Thiscyclization reaction can be performed in a non-protic solvent such asmethylene chloride or chloroform to give the intermediate of the formula##STR48##

The compounds of formula XXIV after removal of the P₂ protecting groupand prior to cyclization wherein X is O can be converted to thecorresponding compounds wherein X is S. This can be done by variousmethods. For example, the compound of formula XXIV after removal of theP₂ group can be treated with triphenylphosphine, diisopropylazodicarboxylate and thioacetic acid. The resulting thioacetate is thentreated with sodium methoxide in methanol to give the correspondingmercaptan which can then be cyclized as described above.

In another method, the compound of formula XXIV after removal of the P₂group is treated by known methods to give the compound of the formula##STR49## wherein L is a leaving group such as methanesulfonyloxy,para-toluenesulfonyloxy, iodo, or bromo. For example, treatment of thecompound of formula XXIV after removal of the P₂ protecting group withmethanesulfonyl chloride gives the compound of formula XXVI wherein L ismethanesulfonyloxy.

The compound of formula XXVI is then treated with cesium thioacetate togive the correspopnding thioacetate. Treatment with sodium methoxide inmethanol gives the corresponding mercaptan which can then be cyclized asdescribed above.

Alternatively, the compound of formula XXIV wherein X is O can beconverted directly to the intermediate of formula XXV by treatment witha strong acid such as trifluoroacetic acid, paratoluenesulfonic acid, ora commercially available polystyrene sulfonate polymer type ion exchangeresin such as Amberlyst 15® in a suitable solvent such as methylenechloride or chloroform.

The N-protecting group is then removed from the compound of formula XXV,for example, by treatment with hydrazine monohydrate when P₁ togetherwith N atom forms a phthalimido group or by treatment withiodotrimethylsilane or palladium on carbon and ammonium formate orhydrogen when P₁ is benzyloxycarbonyl or by treatment with hydrochloricacid in dioxane or other strong acid when P₁ is t-butoxycarbonyl to givethe fused bicyclic ring compound of formula III.

In still another method when Y is CH₂, the N-protected amino acid offormula XXII can be coupled with the hydroxy amino acid ester of theformula ##STR50## to give the dipeptide of the formula ##STR51## whereinP₁ and P₂ are as defined above. This coupling reaction is preferablyperformed in the presence of a coupling reagent such asmethanesulfonyloxybenzotriazole or ethyl-3-(dimethylamino)propylcarbodiimide.

Hydroxy compound XXVIII is then oxidized to the aldehyde of the formula##STR52## by treating with oxalyl chloride/dimethylsulfoxide followed bya tertiary amine in a non-protic solvent such as methylene chloride. Thealdehyde of formula XXIX is then treated as described above to removethe P₂ protecting group and then subjected to an acid catalyzedcyclization reaction as described above to give the intermediate offormula XXV.

The starting material of formula XXIII wherein m is one can be preparedby selective protection of the N-atom of L-s-hydroxynorleucine to give##STR53## wherein P₃ is an N-protecting group. For example, P₃ and theN-atom can form a phthalimido moiety. The N-protectedL-ε-hydroxynorleucine of formula XXX is then treated to introduce the R₃acid protecting group such as by treatment with methyl iodide in thepresence of base or by treatment with a strong acid in methanol whereinR₃ is methyl. This ester is then oxidized to give the aldehyde of theformula ##STR54## The aldehyde of formula XXXI is then treated with theorthoformate of the formula ##STR55## in the presence of a strong acidcatalyst and the corresponding alcohol, i.e. HO-alkyl wherein alkyl isthe same as in the orthoformate of formula XXXII, to give ##STR56##Removal of N-protecting group P₃ such as by treatment with hydrazinehydrate when P₃ and the N-atom forms a phthalimido moiety yields thestarting material of formula XXIII wherein m is one.

The starting material of formula XXIII wherein m is zero can be preparedby protecting the N-atom of γ-benzyl glutamate to give ##STR57## whereinP₃ is an N-protecting group such as t-butyloxycarbonyl or where P₃ andthe N-atom can form a phthalimido moiety. The N-protected glutamic acidof formula XXXIV is then treated to introduce the R₃ acid protectinggroup, as described above, to give ##STR58## Hydrogenolysis when R₃ islower alkyl removes the benzyl ester group from compound XXXV to give##STR59## Selective reduction of compound XXXVI such as by treatmentwith ethanethiol, ethyl-3-(3-dimethlamino)propyl carbodiimide, anddimethylaminopyridine followed by triethylsilane, palladium on carbon,and acetonitrile gives the aldehyde of the formula ##STR60## Thealdehyde of formula XXXVII is then treated with the orthoformate offormula XXXII as described above and the N-protecting group P₃ isremoved as described above to give the starting material of formulaXXIII wherein m is zero.

The hydroxy amino acid ester starting material of formula XXVII can beprepared by reacting a solution of diethyl acetamidomalonate with astirred suspension of sodium hydride followed by reaction with ahaloalkylacetate of the formula ##STR61## wherein halo is Br, I, or Clto give the compound of the formula ##STR62##

A solution of the diethyl ester of formula XXXIX is treated with sodiumhydroxide and heat and then acidified and heated again to give thehydroxy amino acid of the formula ##STR63##

The hydroxy amino acid of formula XL is then treated with porcine kidneyacylase or other suitable hydrolyzing enzyme to give the resolvedhydroxy amino acid of the formula ##STR64##

The hydroxy amino acid of formula XLI is then converted to the ester offormula XXVII by conventional means. For example, the hydroxy amino acidof formula XLI can be treated in methanol with trimethylsilyl chlorideto give the hydrochloride salt of the methyl ester of formula XXVII.

The starting materials of formula XXII can be prepared as follows. WhenX is O, the hydroxy α-amino acid of the formula ##STR65## is reacted tointroduce the P₁ and P₂ protecting groups. For example, treatment of theacid of formula XLII with N-carbethoxyphthalimide in the presence ofsodium carbonate followed by treatment with chlorotriphenylmethane andtriethylamine gives the starting material of formula XXII wherein X isO, P₁ together with N-atom forms a phthalimido, and P₂ is trityl.Alternatively, treatment of the acid of formula XLII withN-(benzyloxycarbonyloxy)succinimide in aqueous sodium carbonate andacetone followed by treatment with t-butyldimethylsilyl chloride or anacylating agent of formula V or VI gives the starting material offormula XXII wherein X is O, P₁ is benzyloxycarbonyl, and P₂ ist-butyldimethylsilyl or an acyl group such as acetyl.

When X is S and n is one, N,N'-bis[(phenylmethoxy)carbonyl]-L-cystinecan be treated with zinc dust and aqueous sulfuric acid to give themercaptan of the formula ##STR66##

The mercaotan of formula XLIII is then treated to introduce the P₂protecting group. For example, treatment of the mercaptan of formulaXLIII with acetic anhydride gives the starting material of formula XXIIwherein X is S, n is one, P₂ is acetyl, and P₁ is benzyloxycarbonyl.

When X is S and n is two, L-methionine can be protected on the N-atom.For example, reaction with benzyl chloroformate orN-(benzyloxycarbonyloxy)succinimide givesN-[(phenylmethoxy)carbonyl]-L-methionine which is then esterified bytreatment with an alcohol, alkyl-OH, in the presence of an acidcatalyst. such as p-toluenesulfonic acid. Treatment with an oxidizingagent such as N-chlorosuccinimide in aqueous solvent gives the sulfoxideof the formula ##STR67##

The sulfoxide of formula XLIV is then treated with an acid anhydridesuch as acetic anhydride to give the compound of the formula ##STR68##Treatment with alkali metal hydroxide, followed by the removal offormaldehyde such as by treatment with a reducing agent, for example,sodium borohydride, followed by treatment with an acid anhydride such asacetic anhydride gives the starting material of formula XXII wherein Xis S, n is two, P₂ is acetyl, and P₁ is benzyloxycarbonyl.

The fused bicyclic ring compounds of formula III wherein Y is S or O andm is one can be prepared by coupling the N-protected amino acid offormula XXII with the amino acid ester of the formula ##STR69## to givethe dipeptide of the formula ##STR70## wherein P₁ and P₂ are as definedpreviously and R₃ is an acid protecting group. This coupling reaction ispreferably performed in the presence of a coupling reagent such asbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphateor ethyl-3-(3-dimethylamino)propyl carbodiimide.

The P₂ protecting group can be selectively removed from the intermediateof formula XLVII such as by treatment with sodium methoxide in methanolwhen P₂ is an acyl group such as acetyl or benzoyl and treatment with anacid such as p-toluenesulfonic acid in methanol when P₂ is a trityl,tetrahydropyranyl, or a hindered silyl group. The resulting product isthen subjected to an acid catalyzed cyclization reaction as describedabove to give the intermediate of the formula ##STR71##

The intermediate of formula XLVIII wherein X is S and n is two can alsobe prepared by treating the compound of formula XLVII wherein X is O andn is two to selectively remove the P₂ group and convert the hydroxy to amercaptan as described above followed by acid catalyzed cyclization.

The N-protecting group is then removed from the compound of formulaXLVIII for example, by treatment with hydrazine monohydrate when P₁together with N atom forms a phthalimido group or by treatment withiodotrimethylsilane or palladium on carbon and ammonium formate orhydrogen when P₁ is benzyloxycarbonyl to give the fused bicyclic ringcompounds of formula III.

The starting material of formula XLVI wherein Y is O can be prepared byreacting the N-phthalimino protected amino acid ester of the formula##STR72## with allyl trichloroacetimidate in the presence oftrifluoromethanesulfonic acid to give the compound of the formula##STR73##

Treatment of the compound of formula L with ozone in methanol thendimethylsulfide followed by the orthoformate of formula XXXII in thepresence of p-toluenesulfonic acid yields the protected compound of theformula ##STR74## Removal of the N-protecting group such as by treatmentwith hydrazine hydrate yields the starting material of formula XLVIwherein Y is O.

The starting material of formula XLVI wherein Y is S can be prepared byreacting the cysteine ester of the formula ##STR75## with thebromoacetal of the formula ##STR76## in the presence of sodium hydrideand potassium iodide to give the amino acid ester of the formula##STR77##

The compounds of formula I contain three asymmetric centers in the fusedbicyclic portion of the structure with additional centers possible inthe side chain. While the optically pure form of the fused bicyclicproducts described above is preferred, all such forms are within thescope of this invention. The above described processes can utilizeracemates, enantiomers, or diastereomers as starting materials. whendiastereomeric compounds are prepared, they can be separated byconventional chromatographic or fractional crystallization methods.Preferably, the hydrogen attached to the bridgehead carbon is in theorientation shown below ##STR78##

The compounds of formula I wherein R₃, R₅ and/or R₇ are hydrogen can beisolated in the form of a pharmaceutically acceptable salt. Suitablesalts for this purpose are alkali metal salts such as sodium andpotassium, alkaline earth metal salts such as calcium and magnesium,salts derived from amino acids such as arginine, lysine, etc. and saltsderived from amines such as alkylamines, e.g. t-butylamine, t-amylamine,etc., substituted alkylamines, e.g. benzylamine, dialkylamines,substituted dialkylamines, e.g. N-methyl glucamine, trialkylamines,substituted trialkylamines, and quaternary ammonium salts. These saltscan be obtained by reacting the acid form of the compound with a basesupplying the desired ion in a medium in which the salt precipitates orin aqueous medium and then lyophilizing.

Preferred compounds of this invention are those wherein:

A is ##STR79## R₂ is hydrogen, ##STR80## or R₁₁ --S--; R₃ is hydrogen orlower alkyl of 1 to 4 carbons;

r is zero or one;

R₁₁ is lower alkyl of 1 to 4 carbons;

R₁ is aryl-CH₂ --, substituted aryl-CH₂ --, heteroaryl-CH₂ --,cycloalkyl-CH₂ -- wherein the cycloalkyl is of 3 to 7 carbons, orstraight or branched chain alkyl of 1 to 7 carbons and R₁₂ is hydrogen;or R₁ and R₁₂ taken together with the carbon to which they are attachedcomplete a cycloalkyl ring of 5 to 7 carbons;

R₆ is lower alkyl of 1 to 4 carbons or phenyl;

n is one or two;

m is zero or one;

X is O or S; and

Y is CH₂, O, or S provided that Y is O or S only when m is one.

Most preferred are the above compounds wherein:

R₂ is hydrogen or ##STR81## especially hydrogen; R₃ is hydrogen;

r is zero or one; especially one;

R₁ is benzyl, cyclopropylmethyl, or straight or branched chain alkyl of3 to 5 carbons, especially benzyl;

R₁₂ is hydrogen;

n is one or two;

m is zero or one;

X is O or S; and

Y is CH₂, O, or S provided that Y is O or S only when m is one.

The single most preferred compound isS-[4α(R*),7α,10aβ]]-octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid, i.e. the compound of the formula ##STR82##

The compounds of formula I wherein A is ##STR83## are dual inhibitorspossessing the ability to inhibit angiotensin converting enzyme andneutral endopeptidase. The compounds of formula I wherein A is ##STR84##are selective inhibitors possessing the ability to inhibit theangiotensin converting enzyme. Thus, the compounds of formula Iincluding their pharmaceutically acceptable salts are useful in thetreatment of physiological conditions in which angiotensin convertingenzyme inhibitors have been shown to be useful. Such conditions includedisease states characterized by abnormalities in blood pressure,intraocular pressure, and renin including cardiovascular diseasesparticularly hypertension and congestive heart failure, glaucoma, andrenal diseases such as renal failure, diabetic nephropathy, and renalimpairment following treatment with cyclosporine or otherimmunosuppressants. Other conditions in which angiotensin convertingenzyme inhibitors have been reported to be useful include hepaticcirrhosis, inhibiting the progression of atherosclerosis, preventing ortreating hypertensive or diabetic retinopathy, improving myocardialdysfunction during or following a myocardial infarction, and preventingrestinosis after angioplasty. The dual inhibitors are also useful in thetreatment of physiological conditions in which neutral endopeptidaseinhibitors have been shown to be useful. Such conditions also includecardiovascular diseases particularly hypertension, hyperaldosteronemia,renal diseases, glaucoma, as well as the relief of acute or chronicpain. Thus, the compounds of formula I are useful in reducing bloodpressure and the dual inhibitors of formula I are additionally usefulfor this purpose due to their diuresis and natriuresis properties. Thedual inhibitors are particularly useful in the treatment of congestiveheart failure.

The compounds of formula I inclduing pharmaceutically acceptable saltsthereof can be administered for these effects in amounts similar tothose employed previously for angiotensin converting enzyme inhibitors.For example, the compounds of formula I can be administered to amammalian host such as man at from about 0.1 mg. to about 100 mg. perkg. of body weight per day, preferably from about 0.5 mg. to about 25mg. per kg. of body weight per day. The compounds of formula I arepreferably administered orally but parenteral routes such assubcutaneous, intramuscular, and intravenous can also be employed as cantopical routes of administration. The daily dose can be administeredsingly or can be divided into two to four doses administered throughoutthe day.

The inhibitors of formula I can be administered in combination withhuman ANF 99-126. Such combination would contain the inhibitor offormula I at from about 1 to about 100 mg. per kg. of body weight andthe human ANF 99-126 at from about 0.001 to about 0.1 mg. per kg. ofbody weight.

The inhibitors of formula I can be administered in combination withother classes of pharmaceutically active compounds. For example, adiuretic, a calcium channel blocker, a potassium channel activator, acholesterol reducing agent, a β-blocker, etc.

The inhibitors of formula I or a pharmaceutically acceptable saltthereof and other pharmaceutically acceptable ingredients can beformulated for the above described pharmacetical uses. Suitablecompositions for oral administration include tablets, capsules, andelixirs, and suitable compositions for parenteral administration includesterile solutions and suspensions. Suitable compositions for treatingglaucoma also include topical compositions such as solutions, ointments,and solid inserts as described in U.S. Pat. No. 4,442,089. About 10 to500 mg. of active ingredient is compounded with physiologicallyacceptable vehicle, carrier, excipient, binder, preservative,stabilizer, flavoring, etc., in a unit dose form as called for byaccepted pharmaceutical practice.

The following examples are illustrative of the invention. Temperaturesare given in degrees centigrade. Thin layer chromatography (TLC) wasperformed in silica gel unless otherwise stated.

EXAMPLE 1[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo7H-pyrido[2,1-b][1,3]-oxazepine-7-carboxylicacid

a) (S)-2-Phthalimido-4-hydroxybutanoic acid, triethylamine salt

A solution of L-homoserine (3.0 g., 25.2 mmol.) and sodium carbonate(2.670 g., 25.2 mmol.) in water (60 ml.) was treated withN-carbethoxyphthalimide (5.570 g., 25.4 mmol.). After stirring at roomtemperature for 2 hours, the solution was acidified with 6N hydrochloricacid and extracted into ethyl acetate. The ethyl acetate extract waswashed with brine, dried (sodium sulfate), and filtered into a solutionof triethylamine (4.0 ml.) in methylene chloride (40 ml.). The cloudysolution was concentrated and triturated with ethyl acetate and ethylether to afford 5.11 g. of the title compound as a white solid; m.p.142°-144° C. TLC (5% acetic acid in ethyl acetate) R_(f) =0.36;

[α]_(D) =-6.2° (c=0.8, chloroform).

Anal. calc'd. for C₁₈ H₂₆ N₂ O₅ : C 61.70; H 7.48; N 7.99 Found: C61.45; H 7.47; N 7.84.

b) (S)-2-Phthalimido-4-(triphenylmethoxy)butanoic acid, triethylaminesalt

A homogeneous solution of the product from part (a) (1.890 g., 5.4mmol.) in chloroform (20 ml.) was treated with triethylamine (80 μl.)followed by solid chlorotriphenylmethane (1.590 g., 5.70 mmol.). Afterstirring at room temperature for 2.5 hours, the solution was partitionedbetween ethyl acetate and 0.1N hydrochloric acid (150 ml.). The organiclayer was washed with water and brine, then dried (sodium sulfate) andfiltered into a solution of triethylamine (1.0 ml.) in methylenechloride (30 ml.). The solution was concentrated to an oil, redissolvedin a small amount of methylene chloride and ethyl acetate and trituratedwith ethyl ether until the solution became turbid. The mixture wasseeded and let stand at room temperature. The resulting precipitate wascollected by filtration, washed with ethyl acetate and ethyl ether, anddried in vacuo to afford 2.538 g. of the title compound as a whitesolid; m.p.=165°-170° C. (decomp.). TLC (10% methanol in chloroform)R_(f) =0.23; [α]_(D) =+7.0° (c=1.2, chloroform).

c) (S)-2-Phthalimido-6-hydroxyhexanoic acid

A solution of (+)-L-ε-hydroxynorleucine [prepared according to theprocedure of Bodanszky et al., J. Med Chem., 1978, 21, 1030-1035] (1.030g., 7.0 mmol.) and sodium carbonate (745 mg., 7.0 mmol.) in water (12ml.) was treated with N-carbethoxyphthalimide (1.495 g., 7.0 mmol.) andthe mixture was stirred at room temperature for 2 hours. The solutionwas filtered, cooled to 0° C., and acidified with 6N hydrochloric acidto afford a white precipitate. The solid was collected by filtration anddried in vacuo at 80° C. for one hour to give 1.297 g. of the titlecompound; m.p. 162°-163° C.; [α]_(D) =-35.7° (c=1.3, methanol).

d) (S)-2-Phthalimido-6,6-dimethoxyhexanoic acid, methyl ester

A slurry of the product from part (c) (3.752 g., 13.5 mmol.) and cesiumcarbonate (2.178 g., 6.7 mmol.) in dimethylformamide (44 ml.) wastreated with methyl iodide (3.0 ml., 6.84 g., 48.2 mmol.). Afterstirring at room temperature for 2 hours, the mixture was diluted withethyl acetate and washed successively with water containing a smallamount of sodium bisulfite, water, 50% saturated sodium bicarbonate, andbrine, then dried (sodium sulfate), filtered and stripped to give theintermediate ester as a colorless oil (3.825 g.). The oil washomogeneous by TLC (1:1-acetone:hexanes) R_(f) =0.37.

A -78° C. solution of oxalyl chloride (1.37 ml., 2.00 g., 15.7 mmol.) indry methylene chloride (58 ml.) was treated dropwise with a solution ofdry dimethylsulfoxide (2.24 ml., 2.47 g., 31.6 mmol.) in methylenechloride (2 ml.). After 10 minutes, a solution of the abovealcohol-ester (3.825 g., 13.1 mmol.) in methylene chloride (10 ml.) wasadded. After an additional 15 minutes, triethylamine (8.0 ml.) was addedand the mixture was stirred at -78° C. for 5 minutes, then warmed to 0°C. The mixture was diluted with ethyl acetate/ethyl ether and wassubsequently washed with 1N hydrochloric acid, water, and brine, thendried (sodium sulfate), filtered and stripped to give the crude desiredaldehyde. The oil was homogeneous by TLC (1:1-acetone:hexanes) R_(f)=0.48.

A solution of the above aldehyde in methanol (17 ml.) and methylenechloride (17 ml.) was treated with trimethyl orthoformate (1.7 ml.)followed by p-toluenesulfonic acid monohydrate (180 mg.). The mixturewas stirred at room temperature for 1.5 hours, then partitioned betweenethyl acetate and 50% saturated sodium bicarbonate. The organic layerwas washed with water and brine, then dried (sodium sulfate), fliteredand stripped. The residue was flash chromatographed (Merck silica gel,1:1-ethyl acetate:hexanes) and the pure product fractions werecrystallized from ethyl acetate/hexanes to give the analytically puretitle product (3.452 g., first crop and 215 mg., second crop) as whiteneedles; m.p. 69°-70° C. TLC (1:1-ethyl acetate:hexanes) R_(f) =0.35;

[α]_(D) =-27.4° (c=1.5, chloroform).

Anal. calc'd. for C₁₇ H₂₁ NO₆ ; C 60.89; H 6.31; N 4.18 Found C 60.80; H6.32; N 4.16.

e)[S-(R*,R*)]-2-[[2-Phthalimido-4-(triphenylmethoxy)-1-oxobutyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

A slurry of the product from part (d) (2.540 g., 7.57 mmol.) in methanol(18 ml.) was treated with hydrazine monohydrate (378 μl., 390 mg, 7.80mmol.). The mixture became homogeneous within 10 minutes. After stirringat room temperature for 3 days, the resulting slurry was filtered,stripped, slurried in methylene chloride, filtered and stripped again toafford the crude intermediate amine as a colorless oil. Meanwhile asolution of the triethylamine salt product from part (b) (4.622 g., 7.80mmol.) in methylene chloride (50 ml.) at 0° C. was treated withbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(3.519 g., 7.95 mmol.). The mixture was stirred for 35 minutes, thentreated with a solution of the above amine in methylene chloride (15ml.). After 10 minutes at 0° C. and 2 hours at room temperature, thesolution was partitioned between ethyl ether and water. The organiclayer was washed with 50% saturated sodium bicarbonate and brine, thendried (sodium sulfate), filtered and stripped. The residue was flashchromatographed (Merck silica gel, 6:4-ethyl acetate:hexanes) to give3.580 g. of pure title compound as a white foam. TLC (6:4-ethylacetate:hexanes) R_(f) =0.32; [α]_(D) =+26.2° (c=0.6, chloroform).

f)[S-(R*,R*)]-2-[(2-Phthalimido-4-hydroxy-1-oxobutyl)amino]-6,6-dimethoxyhexanoicacid, methyl ester

A solution of the product from part (e) (5.420 g., 8.0 mmol.) inmethanol (60 ml.) was treated with p-toluenesulfonic acid monohydrate(520 mg.). After stirring at room temperature for 1.5 hours, the mixturewas partitioned between ethyl acetate and dilute sodium bicarbonate. Thephases were separated and the aqueous layer was extracted again withethyl acetate. The pooled organic extracts were washed with brine, dried(sodium sulfate), filtered and stripped. The residue was flashchromatographed (Merck silica gel, 8:2-ethyl acetate:hexanes followed by5% methanol in ethyl acetate) to afford 2.860 g. of the title product asa colorless oil. TLC (7:3-ethyl acetate:hexanes) R_(f) =0.26; [α]_(D)=+18.7° (c=1.3, chloroform).

g)[4S-(4α,7α,10aβ)]-Octahydro-4-phthalimido-5-oxo-7H-pyrido[2,1-b][1,3]oxazepine7-carboxylicacid, methyl ester

A solution of the product from part (f) (2.10 g., 4.95 mmol.) inmethylene chloride (100 ml.) was treated with Amberlyst® 15 ion exchangeresin (240 mg., pre-washed successively with 6N hydrochloric acid,water, tetrahydrofuran, then methylene chloride). After stirring at roomtemperature for 2.5 hours, the solution was filtered, stripped and flashchromatographed (Merck silica gel, 6:4-ethyl acetate:hexanes followed by100% ethyl acetate) to give 1.40 g. of title product as a white foam.

h) (s)-2-(Acetylthio)benzenepropanoic acid

Sodium nitrite (10.3 g., 280 mmol.) was added to a solution ofD-phenylalanine (30.0 g., 181 mmol.) and potassium bromide (73.5 g.) insulfuric acid (2.5N, 365 ml.) over a period of one hour whilemaintaining the temperature of the reaction mixture at 0° C. The mixturewas stirred for an additional hour at 0° C. and then for one hour atroom temperature. The reaction solution was extracted with ether, theether was back extracted with water, and the ether layer was dried oversodium sulfate. Ether was removed in vacuo, and distillation of the oilyresidue afforded 25.7 g. of (R)-2-bromo-3-benzenepropanoic acid; b.p.141° C. (0.55 mm. of Hg); [α]D=+14.5° (c=2.4, chloroform).

A mixture of thioacetic acid (7 ml., 97.9 mmol.) and potassium hydroxide(5.48 g., 97.9 mmol.) in acetonitrile (180.5 ml.) was stirred underargon at room temperature for 13/4 hours. The mixture was cooled in anice-bath, and a solution of (R)-2-bromo-3-benzenepropanoic acid (20.4g., 89 mmol.) in acetonitrile (20 ml.) was added over a ten minuteperiod. The reaction was stirred under argon at room temperature for 5hours, filtered, and the acetonitrile was removed in vacuo. The oilyresidue was redissolved in ethyl acetate and washed with 10% potassiumbisulfate and water. Removal of the ethyl acetate in vacuo afforded 19.6g. of crude product. The crude product was purified via itsdicyclohexylamine salt using isopropyl ether as solvent forcrystallization. An analytical sample of(S)-2-(acetylthio)benzenepropanoic acid, dicyclohexylamine salt wasprepared by recrystallization from ethyl acetate; m.p. 146°-147° C.;[α]_(D) =-39.6° C. (c=1.39, chloroform).

Anal. calc'd. for C₁₁ H₁₂ O₃ S•C₁₂ H₂₃ N: C, 68.11; H, 8.70; N,3.45; S,7.91 Found: C, 67.93; H, 8.71; N, 3.37; S, 7.94.

The free acid was regenerated by partitioning the dicyclohexylamine saltbetween 5% potasssium bisulfate and ethyl acetate to yield(S)-2-(acetylthio)benzenepropanoic acid; [α]_(D) =-70.1° C. (c=1.91,chloroform).

Anal. calc'd. for C₁₁ H₁₂ O₃ S: C, 58.91; H, 5.39; S, 14.30 Found: C,58.73; H, 5.41; S, 14.53.

i)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]oxazepine-7-carboxylicacid, methyl ester

The product from part (g) (620 mg., 1.66 mol.) in methanol (10 ml.) wastreated with hydrazine monohydrate (85 μl., 88 mg., 1.75 mol.) and thesolution was stirred at room temperature for 44 hours. The mixture wasfiltered and the solid was washed with methanol. The filtrate wasstripped, triturated with methylene chloride, filtered again andstripped to give the crude amine as a cloudy oil (about 400 mg.).

A cold (0° C.) solution of (S)-2-(acetylthio)benzenepropanoic acid (410mg., 1.83 mmol.) and triethylamine (250 μl., 182 mg., 1.80 mmol.) inmethylene chloride (10 ml.) was treated with the above amine (as asolution in 8 ml. methylene chloride) followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(808 mg, 1.83 mmol.). The clear, nearly colorless solution was stirredat 0° C. for 40 minutes and then at room temperature for 2 hours. Themixture was partitioned between ethyl acetate/ethyl ether and water. Theorganic layer was washed successively with 50% saturated sodiumbicarbonate and brine, then dried (sodium sulfate), filtered andstripped. The residue was flash chromatographed (Merck silica gel,60-70% ethyl acetate in hexanes) to give 602 mg., of pure title productas a white foam; TLC (6:4-ethyl acetate:hexanes) R_(f) =0.27.

j) [4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]oxazepine-7-carboxylicacid

A 0° C. solution of the product from part (i) (590 mg., 1.32 mmol.) inmethanol (10 ml., de-oxygenated via argon bubbling) was treated with 1Nsodium hydroxide (7 ml., de-oxygenated via argon bubbling). Afterstirring for 15 minutes, the solution was warmed to room temperature andstirring under argon was continued for an additional 4.5 hours. Themixture was acidified with 5% potassium bisulfate, diluted with waterand extracted with ethyl acetate. The ethyl acetate extract was washedwith water and brine, then dried (sodium sulfate), filtered andconcentrated to approximately 3 ml. The residue was slurried in ethylacetate and a little hexane and the resulting solid was collected byfiltration and dried in vacuo to give 413 mg. of the title product; m.p.180.5° C. (decomp.). TLC (2% acetic acid in ethyl acetate) R_(f) =0.39;

[α]_(D) =-37.6° (c=0.36, methanol). HPLC: YMC S3 ODS column (6.0×150mm); eluted with 40% A: 90% water-10% methanol-0.2% phosphoric acid and60% B: 10% water-90% methanol-0.2% phosphoric acid; flow rate 1.5 ml/mindetecting at 220 nm; t_(R) =6.73 min (95.7%).

Anal. calc'd. for C₂₀ H₂₈ N₂ O₄ S•0.12 ethyl acetate: C, 58.05; H, 6.24;N, 6.95; S, 7.96 Found C, 58.23; H, 6.34; N, 6.83; S, 7.81.

EXAMPLE 2[3R-[3α(S*),6α,9aβ]]-Hexahydro-3-[(2-mercapto-1-oxo-3-phenylpropy)amino]-4-oxo-2H,6H-pyrido[2,1-b][1,3]-thiazine-6-carboxylicacid

a) N-[(Phenylmethoxy)carbonyl]-L-cysteine

A solution of N,N'-bis[(phenylmethoxy)carbonyl]-L-cystine (4.658 g.,9.16 mmol.) in methanol (35 ml.) was treated with 2N sulfuric acid (23ml.) followed by portionwise treatment with zinc dust (2.442 g., 37.3mmol.). The mixture was heated at 70° C. for 1.5 hours, filtered whilestill warm, and concentrated on the rotovap. The residual solution wasextracted with ethyl ether and the ethereal extract was washed withwater and brine, then dried (sodium sulfate), filtered and stripped. Theresidue (oil) was dissolved in carbon tetrachloride, cooled to 0° C.,and seeded to slowly afford a precipitate. The solid was collected byfiltration and washed with cold carbon tetrachloride to give 2.648 g. ofproduct. The mother liquor was stripped, flash chromatographed (Mercksilica gel, ethyl acetate followed by 4% acetic acid in ethyl acetate)to give additional product after crystallization (246 mg.). The totalyield of product was 2.894 g. TLC (5% acetic acid in ethyl acetate)R_(f) =0.58.

b) S-Acetyl-N-[(Phenylmethoxy)carbonyl]-L-cysteine

A homogeneous solution of the product from part (a) (2.70 g., 10.6mmol.) in water (30 ml., de-oxygenated via argon bubbling ) containingpotassium bicarbonate (2.140 g., 21.4 mmol.) was treated with aceticanhydride (8.0 ml., 8.66 g, 84.8 mmol.). After 10 minutes at roomtemperature, the mixture was acidified with 10% hydrochloric acid andextracted with ethyl ether. The ethyl ether extract was washed twicewith water and brine, then dried (sodium sulfate,), filtered andstripped to give an oil. The residue was azeotroped three times withtoluene and twice with ethyl ether/hexane, after which time the oilcrystallized. The residue was triturated with ethyl ether/hexane and thesolid was collected by filtration to give 2.19 g., of pure titleproduct. TLC (5% acetic acid in ethyl acetate) R_(f) =0.56.

c)(S)-2-[[N-[(Phenylmethoxy)carbonyl-S-acetyl-L-cysteninyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

A slurry of (S)-2-phthalimido-6,6-dimethoxyhexanoic acid, methyl ester[prepared as described in example 1(d), 1.158 g., 3.45 mmol.) inmethanol (12 ml.) was treated with hydrazine monohydrate (176 μl., 182mg., 3.63 mmol.). The mixture became homogeneous within 10 minutes.After stirring at room temperature for 67 hours, the resulting slurrywas filtered, stripped, slurried in methylene chloride, filtered andstripped again to afford the crude intermediate amine as a colorlessoil. Meanwhile a partial slurry of the product from part (b) (1.185 g.,3.98 mmol.) in methylene chloride (14 ml.) was treated withtriethylamine (555 μl., 403 mg., 3.98 mmol.). The now homogeneoussolution was cooled to 0° C., treated with the above amine as a solutionin methylene chloride (7 mL), then treated withbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(1.762 g., 3.98 mmol.). The mixture was stirred at 0° C. for 2.5 hours,then at room temperature for 45 minutes. The solvent was removed and theresidue was partitioned between ethyl acetate and water. The organiclayer was washed with 50% saturated sodium bicarbonate and brine, thendried (sodium sulfate), filtered and stripped. The residue was flashchromatographed (Merck silica gel, 65:35-ethyl acetate:hexanes) to give1.15 g., of the pure title product as a white foam. TLC (75:25-ethylacetate:hexanes) R_(f) =0.42.

Analysis Calc'd. for C₂₂ H₃₂ N₂ O₈ S: C, 54.53; H, 6.66; N, 5.78; S,6.62; Found C, 54.79; H, 6.72; N, 5.77; S, 6.95.

d)[3R-(3α,6α,9aβ)]-Hexahydro-3-[[(phenylmethoxy)carbonyl]amino]-4-oxo-2H,6H-pyrido[2,1-b][1,3]-thiazine-6-carboxylicacid, methyl ester

A de-oxygenated (argon bubbling) solution of the product from part (c)(1.040 g., 2.15 mmol.) in methanol (12 ml.) at 0° C. was treated withsodium methoxide (25% by weight in methanol, 490 μl., 463 mg., 2.14mmol.). After 20 minutes, the mixture was quenched with saturatedammonium chloride, diluted with water, and extracted with ethyl acetate.The ethyl acetate extract was washed with water and brine, then dried(sodium sulfate), filtered and stripped. The residue was redissolved inmethylene chloride (200 ml.) and stirred at room temperature withAmberlyst® 15 ion exchange resin (820 mg., pre-washed successively with6N hydrochloric acid, water, tetrahydrofuran, then methylene chloride).After 3 hours, the solution was filtered, stripped and flashchromatographed (Merck silica gel, 65:35-ethyl acetate:hexanes) to give757 mg. of the title product as a colorless oil. TLC (75:25-ethylacetate:hexanes) R_(f) =0.58.

e) [3R-(3α,6α,9aβ)]-Hexahydro-3-amino-4-oxo-2H,6H-pyrido[2,1-b][1,3]thiazine-6-carboxylic acid, methyl ester

A solution of the product from part (d) (752 mg., 1.99 mmol.) in drymethylene chloride (15 ml.) was treated at room temperature withiodotrimethylsilane (620 μl., 872 mg., 4.36 mmol.). After stirring for 3hours, the mixture was quenched with water, treated with a small amount10% hydrochloric acid, and extracted with ethyl ether. The layers wereseparated and the ethereal layer was back-extracted with water. Thepooled aqueous layers were made basic (pH 13) with 10% sodium hydroxideand extracted twice with methylene chloride. The pooled methylenechloride extracts were dried (sodium sulfate), filtered and stripped togive 290 mg. of crude title product as a colorless oil. TLC (10%methanol in methylene chloride) R_(f) =0.38.

f)[3R-[3α(S*),6α,9aβ]]-Hexahydro-3-[[2-(acetylthio)1-oxo-3-phenylpropyl]amino]-4-oxo-2H,6H-pyrido[2,1-b][1,3]thiazine-6-carboxylicacid, methyl ester

A cold (0° C.) solution of (S)-2-(acetylthio)benzenepropanoic acid (294mg., 1.31 mmol.) and triethylamine (180 μL, 131 mg., 1.29 mmol.) inmethylene chloride (8 ml.) was treated with the product from part (e)(287 mg., 1.17 mmol.) as a solution in 6 ml. methylene chloride.Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(575 mg., 1.30 mmol.) was then added. The clear, nearly colorlesssolution was stirred at 0° C. for 1 hour and then at room temperaturefor 1 hour. The solvent was removed by rotary evaporation and theresidue was partitioned between ethyl acetate and 5% potassiumbisulfate. The organic layer was washed successively with water, 50%saturated sodium bicarbonate and brine, then dried (sodium sulfate),filtered and stripped. The residue was flash chromatographed (Mercksilica gel, 1:1-ethyl acetate:hexanes) to give 412 mg. of the pure titleproduct as a white foam. TLC (1:1-ethyl acetate:hexanes) R_(f) =0.27;[α]_(D) =-107.0° C.(c=0.6, chloroform).

g)[3R-[3α(S*),6α,9aβ]]-Hexahydro-3-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-4-oxo-2H,6H-pyrido-2,1-b][1,3]thiazine-6-carboxylicacid

A 0° C. solution of the product from part (f) (406 mg, 0.90 mmol) inmethanol (5 ml., de-oxygenated via argon bubbling) was treated with 1Nsodium hydroxide (5 ml., deoxygenated via argon bubbling). Afterstirring for one hour, the solution was warmed to room temperature andstirring under argon was continued for an additional 1.25 hours. Themixture was acidified with 5% potassium bisulfate, diluted with waterand extracted with ethyl acetate. The ethyl acetate extract was washedwith water and brine, then dried (sodium sulfate), filtered andstripped. The residue was flash chromatographed twice (Merck silica gel,2% acetic acid in ethyl acetate). Product fractions were checked byHPLC. The desired fractions were pooled, stripped, and azeotroped twicewith ethyl acetate. The residue was taken up in a small amount of ethylacetate and triturated with hexanes. The solvent was stripped and theresidue was slurried in hexanes, stripped and dried in vacuo to give98.3 mg. of the title product as a hard white foam. TLC(2% acetic acidin ethyl acetate) R_(f) =0.46; [α]_(D) =-57.0° (c=0.4, chloroform).HPLC: YMC S3 ODS column (6.0×150 mm); eluted with 40% A: 90% water-10%methanol-0.2% phosphoric acid and 60% B: 10% water-90% methanol-0.2%phosphoric acid; flow rate 1.5 mL/min detecting at 220 nm; t_(R) =8.33min.(95.0%).

Anal. calc'd. for C₁₈ H₂₂ N₂ O₄ S₂ •0.2 ethyl acetate: C, 54.79; H,5.77; N, 6.80; S, 15.56. Found C, 54.59; H, 6.04; N, 6.59; S, 15.16.

EXAMPLE 3[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylicacid

a)[S-(R*,R*)]-2-[[2-Phthalimido-4-(acetylthio)-1-oxobutyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

A cold (0° C.) solution of triphenylphosphine (1.143 g., 4.36 mmol.) intetrahydrofuran (20 ml.) was treated with diisopropyl azidodicarboxylate(860 μl., 883 mg., 4.37 mmol.). Within 5 minutes a white slurrydeveloped. After 30 minutes, a solution of[S-(R*,R*)-2-[(2-phthalimido-4-hydroxy-1-oxobutyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester [prepared as described in Example 1(f), 928 mg., 2.19mmol.] in tetrahydrofuran (8 ml.) was added followed by neat thioaceticacid (312 μl., 332 mg., 4.36 mmol.). The mixture was stirred at 0° C.for 1.25 hours, then partitioned between 50% saturated sodiumbicarbonate and ethyl acetate. The ethyl acetate extract was washed withbrine, dried (sodium sulfate), filtered and stripped. The residue wasredissolved in ethyl acetate and treated with a small amount of hexaneto precipitate triphenylphosphine oxide. The mixture was filtered andthe filtrate was flash chromatographed (Merck silica gel, 65:35-ethylacetate:hexanes) to give 894 mg. of the title product as a colorlessoil. TLC (75:25-ethyl acetate:hexanes) R_(f) =0.43.

b)[4S-(4α,7α,10aβ)]-Octahydro-4-phthalimido-5-oxo-7H-pyido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A de-oxygenated (argon bubbling) solution of the product from part (a)(814 mg., 1.65 mmol.) in methanol (15 ml.) at 0° C. was treated withsodium methoxide (25% by weight in methanol, 1.05 ml., 4.6 mmol.). After5 minutes, the mixture was quenched with saturated ammonium chloride,diluted with water, and extracted with ethyl acetate. The ethyl acetateextract was washed with water and brine, then dried (sodium sulfate),filtered and stripped. The residue was redissolved in methylene chloride(180 ml.) and stirred at room temperature with Amberlyst® 15 ionexchange resin (285 mg., pre-washed successively with 6N hydrochloricacid. water, tetrahydrofuran, then methylene chloride). After 46 hours,the solution was flitered, stripped and flash chromatographed (Mercksilica gel, 1:1-ethyl acetate:hexanes) to give 314 mg. of the titleproduct as a white foam. Trituration of the foam with ethyl etherproduced the title product as a white solid; m.p.=147°-148° C. TLC(75:25-ethyl acetate:hexanes) R_(f) =0.56; [α]_(D) =-143.2° (c=0.6,chloroform).

c)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

The product from part (b) (280 mg., 0.72 mmol.) in methanol (8 ml.) wastreated with hydrazine monohydrate (42 μl., 43.3 mg., 0.86 mmol.) andthe solution was stirred at room temperature for 67 hours. The mixturewas filtered and the solid was washed with methanol. The filtrate wasstripped, triturated with methylene chloride, filtered again andstripped to give the crude amine as a yellow oil (about 205 mg.).

A cold (0° C.) solution of (S)-2-(acetylthio)benzenepropanoic acid (178mg., 0.79 mmol.) and triethylamine (111 μl., 80 mg., 0.80 mmol.) inmethylene chloride (3 ml.) was treated with the above amine (as asolution in 7 ml. methylene chloride) followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(353 mg., 0.80 mmol.). The solution was stirred at 0° C. for 1 hour andthen at room temperature for 2 hours. The solvent was stripped and theresidue was partitioned between ethyl acetate and 5% potassiumbisulfate. The organic layer was washed successively with water, 50%saturated sodium bicarbonate and brine, then dried (sodium sulfate),filtered and stripped. The residue was flash chromatographed (Mercksilica gel, 1:1-ethyl acetate:hexanes) to give 272 mg. of the pure titleproduct as a white foam.

d)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

A room temperature solution of the product from part (c) (227 mg., 0.49mmol.) in methanol (5 ml., de-oxygenated via argon bubbling) was treatedwith 1N sodium hydroxide (8 ml,, deoxygenated via argon bubbling). Afterstirring for 1 hour, the mixture was acidified with 10% hydrochloricacid, diluted with water and extracted with ethyl acetate. The ethylacetate extract was washed with water and brine, then dried (sodiumsulfate), filtered and concentrated. The resulting solid was slurried inethyl acetate and collected by filtration. The filtrate was flashchromatographed (Merck silica gel, 1% acetic acid in ethyl acetate) andthe desired fractions were pooled, stripped, and triturated with ethylacetate/ethyl ether to give additional solid. The solids were pooled togive a total of 150 mg. of the title product; m.p. 216°-217° C.(decomp.). TLC (2% acetic acid in ethyl acetate) R_(f) =0.56;

[α]_(D) =-72.6° (c=0.28, dimethylformamide). HPLC YMC S3 ODS column(6.0×150 mm); eluted with 40% A: 90% water-10% methanol-0.2% phosphoricacid and 60% B: 10% water-90% methanol-0.2% phosphoric acid; flow rate1.5 ml/min detecting at 220 nm; t_(R) =9.48 min. (97.4%).

Anal. calc'd. for C₁₉ H₂₄ N₂ O₄ S₂ •0.14 ethyl acetate: C, 55.82; H,6.02; N, 6.66; S, 15.24; Found C, 55.53; H, 6.01; N, 6.63; S, 14.91.

EXAMPLE 4[4S-[4α(R*),7α,9αβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]oxazepine-7-carboxylicacid

a) (S)-2-Phthalimido-5-oxo-5-(phenylmethoxy)pentanoic acid

To a solution of γ-benzyl-L-glutamate (17.49 g., 73.70 mmol.) in aqueous(180 ml.) sodium carbonate (7.81 g., 73.70 mmol.) and dioxane (120 ml.)was added N-carbethoxyphthalimide (16.50 g., 75.27 mmol., 1.02 eq.).After stirring at room temperature for 4.5 hours, The reaction mixturewas acidified with 6N hydrochloric acid (30 ml.) and extracted intoethyl acetate (2×400 ml.). The combined ethyl acetate extracts werewashed with 50% brine (200 ml.), and brine (200 ml.), dried over sodiumsulfate, filtered, concentrated and dried in vacuo to yield a crude oil(41.4 g.). To a solution of the crude residue in ethyl ether (100 ml.)was added dicyclohexylamine (14 ml.). After standing in the refrigeratorovernight, the ethyl ether was removed by rotary evaporation and theoily residue was crystallized from ethyl acetate/hexane. The resultingprecipitate was collected by filtration, washed with hexane and dried invacuo to yield 21.21 g. of the title product as the dicyclohexyl aminesalt. A suspension of this dicyclohexylamine salt in ethyl acetate (200ml.) was washed with 5% potassium bisulfate (3×50 ml.), brine (50 ml.)and dried over magnesium sulfate, filtered and concentrated to yield13.5 g. of the title product as a white foam. TLC: (3% acetic acid in9:1 ethyl acetate:heptane) R_(f) =0.30.

b) (S)-2-Phthalimido-5-oxo-5-(phenylmethoxy)pentanoic acid, methyl ester

To a solution of the product from part (a) (13.22 g., 36.0 mmol.) andcesium carbonate (5.86 g., 18.0 mmol.) in dimethylformamide (100 ml.)was added iodomethane (8.1 ml., 129.6 mmol., 3.6 eq.). The yellowsolution was stirred for 2.5 hours, and was then partitioned betweenethyl acetate (300 ml.) and water (250 ml.). The ethyl acetate extractwas washed with 5% sodium bicarbonate (200 ml.) and brine, dried overmagnesium sulfate, filtered and concentrated to yield 13.68 g. a yellowoil. The residue was purified by chromatography on a 5×20 cm. silica gelcolumn eluting with 30% ethyl acetate/hexane. The desired fractions werecombined and concentrated to yield 10.0 g of the title product. TLC(1:1, ethyl acetate:hexane) R_(f) =0.45.

c) (S)-2-Phthalimido-4-(carboxy)butanoic acid, methyl ester

To a solution of the product from part (b) (10.0 g., 26.22 mmol.) inethyl acetate (115 ml.) was added 20% palladium hydroxide on carboncatalyst (1.90 g.) and the resulting suspension was stirred underhydrogen atmosphere (balloon) for 2.5 hours. The mixture was filtered,washed thoroughly with ethyl acetate, concentrated and dried in vacuo toyield 7.29 g. of crude title product as a white solid; m.p. 137°-138° C.TLC (10% methanol/methylene chloride) R_(f) =0.43.

d) (S)-2-Phthalimido-5-oxo-5-(ethylthio)pentanoic acid, methyl ester

To a solution of the product from part (c) (7.27 g., 24.95 mmol.) inmethylene chloride (125 ml.) at 0©C. under argon was added ethanethiol(4.81 ml., 64.92 mmol., 2.6 eq), 4-dimethylaminopyridine (609 mg., 4.99mmol., 0.2 eq.) and ethyl-3-(3-dimethylamino)propyl carbodiimide,hydrochloride salt (5.27 g., 27.47 mmol., 1.1 eq.). After stirring at 0°C. for 2 hours and at room temperature for 1 hour the reaction wasconcentrated, diluted with ethyl acetate (400 ml.) and washed with 5%potassium bisulfate (200 ml.), saturated sodium bicarbonate (200 ml.),and brine (200 ml.), dried over sodium sulfate, filtered, concentratedand dried in vacuo to yield 8.30 g. of title product as a crude oil. TLC(1:1, ethyl acetate:hexane) R_(f) =0.47.

e) (S)-2-Phthalimido-5-oxopentanoic acid, methyl ester

A suspension of the product from part (d) (8.30 g., 24.75 mmol.) and 10%palladium on carbon (1.24 g.) in acetonitrile (150 ml.) under argon wastreated dropwise with triethylsilane (7.91 ml., 49.5 mmol., 2 eq.).After stirring at room temperature for 45 minutes, the mixture wasfiltered, concentrated and dried in vacuo. The crude residue waspurified by chromatography on a 5×25 cm silica gel column eluting with25% ethyl acetate/hexane (4 l.) followed by 35% ethyl acetate/hexane (2l.). The desired fractions were combined to yield 5.60 g. of titleproduct. TLC (1:1, ethyl acetate:hexane) R_(f) =0.32.

f) (S)-2-Phthalimido-5,5-dimethoxypentanoic acid, methyl ester

A solution of the product from part (e) (5.60 g., 20.34 mmol.) inmethanol (60 ml.) and methylene chloride (40 ml.) was treated withtrimethylorthoformate (3.8 ml., 34.59 mmol., 1.7 eq.) andp-toluenesulfonic acid monohydrate (280 mg.). After stirring at roomtemperature for 1.5 hours the reaction was quenched with 2 ml. ofsaturated sodium bicarbonate, concentrated, and partitioned betweenethyl acetate (400 ml.) and water (100 ml.). The ethyl acetate extractwas washed with saturated sodium bicarbonate (100 ml.), brine (100 ml.),dried over magnesium sulfate, filtered and concentrated to a crude oil.The crude residue was purified by chromatography on a 5×20 cm silica gelcolumn eluting with 30% ethyl aceatate/hexane (2 l.). The desiredfractions were combined, concentrated and dried in vacuo to yield 6.20g. of title product. TLC (1:1, ethyl acetate:hexane) R_(f) =0.40.

g) (S)-2-Amino-5,5-dimethoxypentanoic acid, methyl ester

A solution of the product from part (f) (6.16 g., 19.18 mmol.) inmethanol (125 ml.) was treated with hydrazine monohydrate (0.98 ml.,20.14 mmol., 1.05 eq). After stirring at room temperature for 6 days,the resulting slurry was filtered, concentrated, triturated in methylenechloride, filtered, concentrated and dried in vacuo to afford 3.57 g. oftitle product as a cloudy oil. TLC (10% methanol in methylene chloride)R_(f) =0.41.

h)[S-(R*,R*)]-2-[[2-Phthalimido-4-(triphenylmethoxy)-1-oxobutyl]amino]-5,5-dimethoxypentanoicacid, methyl ester

A solution of (S)-2-phthalimido-4-(triphenylmethoxy)butanoic acid,triethylamine salt [prepared as described in Example 1(b), 11.62 g.,19.60 mmol., 1.05 eq.] in methylene chloride (100 ml.) at 0° C. wastreated with benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate reagent (8.67 g., 19.60 mmol., 1.05 eq.). Themixture was stirred for 45 minutes at 0° C., then treated with asolution of the product from part (g) (3.57 g., 18.67 mmol.) inmethylene chloride (50 ml.). After 10 minutes at 0° C. and 2 hours atroom temperature, the solution was partitioned between ethyl acetate(300 ml.) and water (100 ml.). The ethyl acetate layer was washed with50% saturated sodium bicarbonate (100 ml.) and brine (100 ml.), driedover magnesium sulfate, filtered and concentrated. The residue waspurified by chromatography on a 5×25 cm silica gel column eluting with1:1 ethyl acetate/hexane affording 8.58 g. of title product. TLC (1:1,ethyl acetate:hexane) R_(f) =0.20.

i)[S-(R*,R*)]-2-[(2-Phthalimido-4-hydroxy-1-oxobutyl)amino]-5,5-dimethoxypentanoicacid, methyl ester

A solution of the product from part (h) (8.58 g., 12.91 mmol.) inmethanol (100 ml.) was treated with p-toluensulfonic acid monohydrate(850 mg.). After stirring at room temperature for 3.5 hours, the mixturewas partitioned between ethyl acetate (200 ml.) and 10% saturated sodiumbicarbonate (100 ml.). The phases were separated and the aqueous layerwas extracted again with ethyl acetate (100 ml.). The combined ethylacetate extracts were washed with brine, dried over magnesium sulfate,filtered and concentrated. The residue was purified by chromatography ona 5×20 cm silica gel column eluting with 8:2 ethyl acetate:hexane (1 l.)followed by 1% methanol in ethyl acetate (2 l.). The desired fractionswere combined, concentrated and dried in vacuo to yield 4.33 g. of titleproduct. TLC (8:2 ethyl acetate:hexane) R_(f) =0.21.

j)[4S-(4α,7α,9aβ]-Octahydro-4-phthalimido-5-oxopyrrolo[2,1-b][1,3]oxazepine-7-carboxylicacid, methyl ester

A solution of the product from part (i) (1.89 g., 4.48 mmol.) inmethylene chloride (90 ml.) was treated with Amberlyst® 15 ion exchangeresin (400 mg., pre-washed successively with 6N hydrochloric acid,water, tetrahydrofuran and methylene chloride). After stirring at roomtemperature for 3 hours the solution was filtered, concentrated andflash chromatographed on a 5×15 cm silica gel column eluting with 6:4ethyl acetate:hexane to afford 1.51 g. of title product as a white foam,TLC (8:2 ethyl acetate:hexane) R_(f) =0.32.

k)[4S-(4α,7α,9aβ)]-4-Amino-octahydro-5-oxopyrrolo[2,1-b][oxazepine-7-carboxylicacid, methyl ester

The product from part (j) (764 mg., 2.13 mmol.) in methanol (15 ml,) wastreated with hydrazine monohydrate (109 μl., 2.24 mmol., 1.05 eq.) andthe solution was stirred at room temperature for 4 days. The mixture wasfiltered and the solid was washed with methanol. The filtrate wasconcentrated, triturated with methylene chloride, filtered again andconcentrated. The residue was purified by chromatography on a 2×15 cmsilica gel column eluting with 3% methanol in methylene chloride (3 l.)followed by 10% methanol in methylene chloride (1 l.). The desiredfractions were combined and concentrated to afford 451 mg. of titleproduct as an oil. TLC (10 % methanol in methylene chloride) R_(f)=0.18.

l)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxopyrrolo[2,1-b][1,3]oxazepine-7-carboxylicacid, methyl ester

A suspension of the dicyclohexylamine salt of(S)-2-acetylthio-3-benzenepropanoic acid [prepared as described inExample 1(h), 870 mg., 2.14 mmol., 1.14 eq.] in ethyl acetate (70 ml.)was washed with 5% potassium bisulfate (5×20 ml.), 50% brine (20 ml.),and brine (20 ml.), dried (anhydrous sodium sulfate), filtered,concentrated and dried in vacuo overnight to give(S)-2-(acetylthio)benzenepropanoic acid.

This free acid was dissolved in dry methylene chloride (10 ml.), cooledto 0° C. (ice-salt bath) and treated with triethylamine (298 μl, 2.14mmol.) followed by a solution of the product from part (k) (430 mg.,1.88 mmol.) in methylene chloride (10 ml.) andbenzotriazol-1-yloxytris(dimethylamino) phosphonium hexafluorophosphate(947 mg., 2.14 mmol., 1.14 eq.). The resultant solution was stirred at0° C. for 50 minutes then at room temperature for 3 hours. The reactionmixture was concentrated, diluted with ethyl acetate (150 ml.), washedwith 0.5N hydrochloric acid (50 ml.), water (50 ml.), saturated sodiumbicarbonate (50 ml.), water (50 ml.) and brine (50 ml.), dried(anhydrous magnesium sulfate), filtered, and evaporated to dryness. Thecrude product was adsorbed onto Celite® and chromatographed on a silicagel column (5×10 cm), eluting with 60% ethyl acetate/hexane (3 l). Thedesired fractions were combined and concentrated, affording 779 mg. ofpure title product. TLC (6:4, ethyl acetate:hexane) R_(f) =0.17.

m)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]oxazepine-7-carboxylicacid

A solution of the product from part (1) (754 mg., 1.74 mmol.) inmethanol (15 ml.) was purged with argon for 30 minutes, cooled to 0° C.(ice-salt bath) then treated dropwise with a previously purged (argon,30 minutes/ solution of 1.0N sodium hydroxide (12 ml.) maintaining thebubbling of argon throughout the addition and length of the reaction.The reaction mixture was stirred at 0° C. for 3 hours, acidified at 0°C. with 5% potassium bisulfate to pH 1 then extracted with ethyl acetate(3×100 ml.). The combined organic extracts were washed with 50% brine(100 ml.), brine (100 ml.), dried (anhydrous sodium sulfate), filtered,evaporated to dryness and dried in vacuo to yield a whine foam. Theresidue was purified by chromatography on a 2.5×15 cm silica gel columneluting with ethyl acetate (500 ml.) and 0.3% acetic acid in ethylacetate (1 l.). The desired fractions were concentrated, stripped withchloroform and dried in vacuo overnight at 50° C. over phosphoruspentoxide to yield the title product as a white foam; m.p. 88°-92° C.;[α]_(D) =-63.8° (c=1.0, methanol). TLC (1% acetic acid in ethyl acetate)R_(f) =0.24.

¹ H-NMR: 400 MHz; CDCl₃ : δ1.80-2.31 (m's, 7H), 3.10 (m, 1H), 3.27 (m,1H), 3.63 (m, 1H), 4.0 (m, 1H), 4.20 (m, 1H), 4.49 (m, 1H), 4.75 (m,1H), 5.23 (m, 1H), 7.19-7.30 (m's, 5H), 7.52 (d, 1H, J=6 Hz).

¹³ C-NMR: 100 MHz; CDCl₃ : δ26.4, 32.0, 32.6, 41.2, 44.2, 53.0, 59.4,70.6, 89.47, 126.9, 128.4, 129.3, 137.4, 171.2, 171.6, 174.8.

Anal. calc'd. for C₁₈ H₂₂ N₂ O₅ S•0.85 H₂ O: C, 54.91; H, 6.07; N, 7.12;S, 8.14 Found: C, 54.85; H, 5.68; N, 7.18, S, 8.14. HPLC: t_(R) =13.5min (96.7%, UV 220); YMC S-3 ODS (C-18) 6.0×150 mm: 30% B:A-100% B:A, 25minute Linear gradient (A=90% water/methanol+0.2% phosphoric acid B=90%methanol/water +0.2% phosphoric acid) flow rate at 1.5 ml./min.

EXAMPLE 5[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

a)[S-(R*,R*)]-2-[[2-Phthalimido-4-(aceylthio)-1-oxobutyl]amino]-5,5-dimethoxypentanoicacid, methyl ester

A 0° C. solution of triphenylphosphine (1.26 g., 4.79 mmol., 1.5 eq.) indry tetrahydrofuran (15 ml.) was treated with diisopropylazodicarboxylate (943 μl., 4.79 mmol.). The resultant white slurry wasstirred for 30 minutes and then treated with a solution of[S-(R*,R*)]-2-[(2-phthalimido-4-hydroxy-1-oxobutyl)amino]-5,5-dimethoxypentanoicacid, methyl ester [prepared as described in Example 4(i), 1.35 g., 3.20mmol.] in dry tetrahydrofuran (15 ml.) followed by neat thiolacetic acid(343 μl., 4.79 mmol.). The mixture was stirred at 0° C. for 1.5 hoursand then partitioned between ethyl acetate (150 ml.) and 50% sodiumbicarbonate (100 ml.). The ethyl acetate layer was washed with brine,dried over magnesium sulfate, filtered, concentrated, adsorbed ontoCelite® and dried in vacuo. The crude material was purified bychromatography on a 2.5×15 cm silica gel column eluting with 1:1 ethylacetate:hexane (1 l.) and 6:4 ethyl acetate:hexane (1 l.). The desiredfractions were combined, concentrated and dried in vacuo affording 1.35g. of title produce as an oil. TLC (8:2, ethyl acetate:hexane) R_(f)=0.42.

b)[S-(R*,R*)]-2-[(2-Phthalimido-4-mercapto-1-oxobutyl)amino]-5,5-dimethoxypentanoicacid, methyl ester

A de-oxygenated (argon bubbling) solution of the product from part (a)(1.33 g., 2.76 mmol.) in methanol (25 ml.) at 0° C. was treated withsodium methoxide (25% by weight in methanol, 1.52 ml., 6.63 mmol., 2.4eq.). After 3 minutes, the mixture was quenched with saturated ammoniumchloride (3 ml.), diluted with water, and extracted with ethyl acetate(100 ml.). The ethyl acetate extract was washed with water (50 ml.) andbrine (50 ml.), dried over sodium sulfate, filtered and concentrated.The residue was purified by chromatography on a 5×15 cm silica gelcolumn eluting with 1:1 (3 l) followed by 8:2 (2 l) ethylacetate:hexane. The desired product containing fractions were combinedand concentrated to yield 853 mg. of title compound as an oil. TLC (8:2,ethyl acetate:hexane) R_(f) =0.43.

c)[4S-(4α,7α,9aβ]]-Octahydro-4-phthalimido-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of product from part (b) (847 mg., 1.93 mmol.) in methylenechloride (20 ml.) was treated with Amberlyst® 15 ion exchange resin (700mg., pre-washed successively with 6N hydrochloric acid, water,tetrahydrofuran and methylene chloride). After stirring at roomtemperature for 17 hours the solution was filtered, concentrated andflash chromatographed on a 2.5×15 cm silica gel column eluting with 1:1ethyl acetate:hexane to afford 691 mg. of title product as a white foam.TLC (8:2 ethyl acetate:hexane) R_(f) =0.48.

d)[4S-(4α,7α,9aβ]-4-Amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

The product from part (c) (899 mg., 2.40 mmol.) in methanol (17 ml.) wastreated with hydrazine monohydrate (122 μl., 2.52 mmol., 1.05 eq.) andthe solution was stirred at room temperature for days. The mixture wasfiltered and the solid was washed with methanol. The filtrate wasconcentrated, triturated with methylene chloride, filtered again,concentrated, and dried in vacuo to yield 572 mg. of title product as acloudy oil. TLC (10% methanol in methylene chloride) R_(f) =0.13.

e)[4S-[4α(R*),7α9aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A suspension of the dicyclohexlamine salt of (S)-2-(acetylthio)benzenepropanoic acid [prepared as described in Example 1(h), 1.045 g.,2.58 mmol., 1.1 eq] in ethyl acetate (100 ml.) was washed with 5%potassium bisulfate (5×25 ml.), 50% brine (25 ml.), and brine (25 ml.),dried (anhydrous sodium sulfate), filtered, concentrated and dried invacuo for one hour to give (S)-2-(acetylthio)-benzenepropanoic acid.

This free acid was dissolved in dry methylene chloride (10 ml.), cooledto 0° C. (ice-salt bath) and treated with triethylamine (360 μl., 2.58mmol.), benzotriazol-1-yloxtris (dimethylamino)phosphoniumhexafluorophosphate (1.141 g., 2.58 mmol.) and then a solution of theproduct from part (d) (572 mg., 2.34 mmol.) in methylene chloride (10ml.). The resultant solution was stirred at 0° C. or 30 minutes then atroom temperature for 2.5 hours. The reaction mixture was concentrated,diluted with ethyl acetate (100 ml.) washed with 0.5N hydrochloric acid(50 ml.), water (50 ml.), saturated sodium bicarbonate (50 ml.), water(50 ml.) and brine (50 ml.), dried (anhydrous magnesium sulfate),filtered, and evaporated to dryness. The crude product was adsorbed ontoCelite® and chromatographed on a silica gel column (5×10 cm), elutingwith 25% (5 l.), 30% (2 l.), 35% (2 l.), and 40% (2 l.) ethylacetate/hexane. The mixed fractions were combined and rechromatographedeluting with the same gradient. The desired fractions were combined andconcentrated, affording 490 mg. of pure title product. TLC (1:1, ethylacetate:hexane) R_(f) =0.16.

f)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (e) (490 mg., 1.09 mmol.) inmethanol:tetrahydrofuran (8 ml.: 4 ml.) was purged with argon for 30minutes, cooled to 0° C. (ice-salt bath) then treated dropwise with apreviously purged (argon, 30 minutes) solution of 1.0N sodium hydroxide(10 ml.) maintaining the bubbling of argon throughout the addition andlength of the reaction. The reaction mixture was stirred at 0° C. for 3hours, acidified at 0° C. with 5% potassium bisulfate to pH 2 thenextracted with ethyl acetate (3×75 ml.). The combined organic extractswere washed with brine (75 ml.), dried (anhydrous sodium sulfate),filtered, evaporated to dryness and dried in vacuo to yield a white foam(489 mg.). The residue was purified by chromatography on a 2.5×15 cmsilica gel column eluting with 9:1 ethyl acetate:heptane (400 ml.) and0.5% acetic acid in 9:1 ethyl acetate:heptane (1 l.). The desiredfractions were concentrated, stripped with methylene chloride/heptaneand dried in vacuo to yield 428 mg. of product. The pure material wasrecrystallized from a mixture of ethyl acetate/methanol/hexane. Thecrystals were collected by filtration, washing thoroughly with ethylether, and dried in vacuo overnight at 40° C. over phosphorus pentoxideto yield 305 mg. of title product as white crystals; m.p. 206°-208° C.;[α]_(D) =-96.3° (c=1.0, methanol). TLC (5% acetic acid in 9:1 ethylacetate:heptane) R_(f) =0.29.

¹ H-NMR: 400 MHz; CDCl₃ w/2 drops CD₃ OD: δ1.94(m, 1H), 2.02(d, 1H, J=9Hz), 2.08(m, 1H), 2.20-2.55(m's, 4H), 2.95(m, 1H), 3.08(m, 1H), 3.23 (m,1H), 3.27 (m, 1H), 3.59 (m, 1H), 4.54 (t, 1H, J =7.3 Hz), 4.60 (m, 1H),5.23 (m, 1H), 7.18-7.3 4 (m's, 5H), 7.63 (d, 1H, J=6 Hz).

¹³ C-NMR: 100 MHz; CDCl₃ w/2 drops CD₃ OD: δ27.6, 31.1, 32.1, 32.9,41.1, 44.0, 52.8, 60.4, 62.2, 126.77, 128.3, 129.0, 137.4, 170.2, 171.4,172.6.

Anal. calc'd. for C₁₈ H₂₂ N₂ O₄ S₂ •0.08 H₂ O: C, 54.60; H, 5.64; N,7.07; S, 16.19 Found: C, 54.65; H, 5.54; N, 7.02, S, 15.80. HPLC: t_(R)=13.0 min (98.8%, UV 220); YMC S-3 ODS (C-18) 6.0×150 mm; 40% B:A-100%B:A, 25 minute linear gradient (A=90% water/methanol+0.2% phosphoricacid; B=90% methanol/water+0.2% phosphoric acid); flow rate at 1.5ml/min.

EXAMPLE 6[4S-[4α(R*),7α9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

The product of Example 5 was also prepared as follows:

a) N-[(Phenylmethoxy)carbonyl]-L-homoserine

N-(Benzyloxycarbonyloxy) succinimide (23.57 g., 94.58 mmol.) was addedto a solution of L-homoserine (10.24 g., 85.98 mmol.) and sodiumbicarbonate (7.95 g., 94.58 mmol., 1.1 eq.) in a mixture of water (100ml.) and acetone (100 ml.). The mixture was stirred at room temperatureovernight. The acetone was removed under reduced pressure (rotovap) andthe aqueous solution was washed with methylene chloride (2×75 ml.). Theaqueous layer was then acidified to pH 2 by addition of 6N hydrochloricacid and extracted with ethyl acetate (2×250 ml.). The combined ethylacetate layers were washed with water (2×100 ml.) and brine, dried oversodium sulfate, filtered, concentrated and dried in vacuo to afford19.54 g. of title product as a white solid. TLC (ethylacetate:n-butanol:acetic acid:water; 2:1:1:1) R_(f) =0.74

b) N-[(Phenylmethoxy)carbonyl]-O-(triphenylmethyl)-L-homoserine

To a suspension of the product from part (a) (19.51 g., 77.04 mmol.) inchloroform (250 ml.) was added triethylamine (12.35 ml., 88.59 mmol.,1.15 eq.). The homogeneous mixture was treated with triphenylmethylchloride (24.70 g., 88.59 mmol.) and the reaction was stirred for 3hours. The reaction mixture was concentrated under reduced pressure(rotovap), partitioned between ethyl acetate (400 ml.) and 5% potassiumbisulfate (200 ml.). The ethyl acetate layer was washed with 5%potassium bisulfate (200 ml.), water (2×200 ml.), and brine (200 ml.),dried over sodium sulfate, filtered, and concentrated to yield 45.4 g.material. The residue was chromatographed on a 10×30 cm silica gelcolumn eluting with 6:4 ethyl acetate:hexane (2 l.) followed by 1%acetic acid in 8:2 ethyl acetate:hexane to give 8.76 g. of pure titlecompound.

c) (S)-2-Amino-5,5-dimethoxypentanoic acid, methyl ester

(S)-2-Phthalimido-5,5-dimethoxypentanoic acid, methyl ester [prepared asdescribed in Example 4(f), 3.35 g., 10.43 mmol.) in methanol (70 ml.)was treated with hydrazine monohydrate (531 μl., 10.95 mmol., 1.05 eq.)and the solution was stirred at room temperature for 6 days. The mixturewas filtered and the solid was washed with methanol. The filtrate wasconcentrated, triturated with methylene chloride, filtered again,concentrated, and dried in vacuo to yield 1.89 g. of title product as acloudy oil. TLC (10% methanol in methylene chloride) R_(f) =0.39.

d)[S-(R*,R*)]-[[2-[[(Phenylmethoxy)carbonyl]amino]-4-(triphenylmethoxy)-1-oxobutyl]amino]-5,5-dimethoxypentanoicacid, methyl ester

A solution of the product from part (b) (5.28 g., 10.65 mmol., 1.1 eq.)in dry methylene chloride (50 ml.) at 0° C. was treated withtriethylamine (1.48 ml., 10.65 mmol.), followed by the product from part(c) (1.85 g., 9.68 mmol.) in dry methylene chloride (30 ml.) andbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(4.71 g., 10.65 mmol., 1.1 eq.). The mixture was stirred for 1 hour at0° C. and then stirred at room temperature for 2 hours. The reactionmixture was partitioned between ethyl acetate (300 ml.) and water (150ml.). The ethyl acetate layer was washed with 50% saturated sodiumbicarbonate (200 ml.) and brine (2×200 ml.), dried over magnesiumsulfate, filtered, concentrated, adsorbed onto Celite® and purified on a7×20 cm silica gel column eluting with 40% ethyl acetate/hexane (3l.),followed by 50% (2 l.) ethyl acetate/hexane affording 4.84 g. of titleproduct. TLC (ethyl acetate: hexane, 1:1) R_(f) =0.22.

e)[S-(R*,R*)]-2-[[2-[[(Phenylmethoxy)carbonyl]amino]-4-hydroxy-1oxobutyl]amino]-5,5-dimethoxypentanoicacid, methyl ester

A solution of the product from part (d) (4.80 g., 7.18 mmol.) inmethanol (70 ml.) was treated with p-toluenesulfonic acid monohydrate(300 mg.). After stirring at room temperature for 2 hours, the mixturewas partitioned between ethyl acetate (400 ml.) and 25% saturated sodiumbicarbonate (200 ml.). The phases were separated and the aqueous layerwas extracted again with ethyl acetate (100 ml.). The combined ethylacetate extracts were washed with brine, dried over magnesium sulfate,filtered and concentrated. The residue was purified by chromatography ona 5×20 cm silica gel column eluting with 7:3 (1 l.), 8:2 (1 l.) ethylacetate:hexane followed by 10% methanol in ethyl acetate (2 l.). Thedesired fractions were combined, concentrated and dried in vacuo toyield 2.92 g. of title product. TLC (ethyl acetate:hexane, 8.2) R_(f)=0.09.

f)[S-(R*,R*)]-2-[[2-[[(Phenylmethoxy)carbonyl]amino]-4-(acetylthio)-1-oxobutyl]amino]-5-5-dimethoxypentanoicacid, methyl ester

A 0° C. solution of triphenylphosphine (3.06 g., 11.65 mmol., 1.7 eq.)in dry tetrahydrofuran (40 ml.) was treated with diisopropylazodicarboxylate (2.29 ml., 11.65 mmol.). The resultant white slurry wasstirred for 30 minutes and then treated with a solution of the productfrom part (e) (2.92 g., 6.85 mmol.) in dry tetrahydrofuran followed byneat thiolacetic acid (833 μl., 11.65 mmol.). The mixture was stirred at0° C. for 2 hours and then partitioned between ethyl acetate (300 ml.)and 50% sodium bicarbonate (200 ml.). The ethyl acetate layer was washedwith brine, dried over magnesium sulfate, filtered, concentrated,adsorbed onto Celite® and dried in vacuo. The crude material waspurified by chromatography on a 5×20 cm silica gel column eluting with1:1 ethyl acetate:hexane (3 l.). The desired fractions were combined,concentrated and dried in vacuo affording 2.58 g. of title product as anoff-white solid. TLC (ethyl acetate:hexane, 8:2) R_(f) =0.40.

g)[S-(R*,R*)]-2-[[2-[[(Phenylmethoxy)carbonyl]amino]-4-mercapto-1-oxobutyl]amino]-5,5-dimethoxypentanoicacid, methyl ester

A de-oxygenated (argon bubbling) solution of the product from part (f)(2.56 g., 5.28 mmol.) in methanol (50 ml.) at 0° C. was treated withsodium methoxide (25% by weight in methanol, 3.62 ml., 15.84 mmol., 3eq.). After 10 minutes, the mixture was quenched with saturated ammoniumchloride (40 ml.), diluted with water (100 ml.), and extracted withethyl acetate (300 ml.). The ethyl acetate extract was washed with water(100 ml.) and brine (150 ml.), dried over magnesium sulfate, filteredand concentrated. The residue was purified by chromatography on a 5×20cm silica gel column eluting with 1:1 (3 l.) ethyl acetate:hexane. Thedesired compound was combined and concentrated to yield 1.99 g. of titleproduct as an oil. TLC (ethyl acetate:hexane, 8:2) R_(f) =0.43.

h)[4S-(4α,7α,9aβ)]-Octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of the product from part (g) (2.16 g., 4.88 mmol.) inmethylene chloride (50 ml.) was treated with Amberlyst® 15 ion exchangeresin (620 mg., pre-washed successively with 6N hydrochloric acid,water, tetrahydrofuran and methylene chloride). After stirring at roomtemperature for 3 hours the solution was filtered concentrated and flashchromatographed on a 5×20 cm silica gel column eluting with 6:4 ethylacetate:hexane to afford 1.34 g. of title product as a white foam. TLC(ethyl acetate:hexane, 8:2) R_(f) =0.51.

i)[4S-(4α,7α,9aβ]-4-Amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of the product from part (h) (1.20 g., 3.17 mmol., strippedwith toluene three times and dried in vacuo overnight] in dry methylenechloride (40 ml.) was treated with iodotrimethylsilane (632 μl., 4.44mmol., 1.4 eq.) and stirred at room temperature under argon for 1.5hours. The mixture was quenched with water (50 ml.), treated with 10%hydrochloric acid (5 ml., pH 1) and washed with ethyl acetate (50 ml.).The aqueous phase was treated with with 10% sodium hydroxide andextracted with methylene chloride (three times). The pooled extractswere dried over sodium sulfate, filtered, concentrated, and dried invacuo to yield a 396 mg. of title product as a clear oil. TLC (10%methanol in methylene chloride) R_(f) =0.10.

j)[4S-[4α(R*),7α,7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid A solution of (S)-2-(acetylthio)benzenepropanoic acid in drymethylene chloride was treated with triethylamine. A solution of theproduct from part (i) in methylene chloride was then added followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate.The resultant solution was worked-up as described in Example 5(e) togive[4S-[4α(R*),7α,9aβ]]-octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester.

A suspension of this methyl ester product in methanol:tetrahydrofuran,purged with argon, was cooled to 0° C. and treated with a previouslypurged solution of 1.0N sodium hydroxide. Work-up as described inExample 5(f) gave the title product.

EXAMPLE 7[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(3-cyclohexyl-2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

a)[4S-(4α,7α,9aβ)]-4-Amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt

A solution of[4S-(4α,7α,9aβ)]-octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester [prepared as described in Example 6(h), 738 mg., 1.95mmol., stripped three times with toluene and dried in vacuo overnight)in dimethylene chloride (25 ml.) was treated with iodotrimethylsilane(389 μl., 2.73 mmol., 1.4 eq.) and stirred at room temperature underargon. After 2 hours the reaction was treated with additional amounts ofiodotrimethyl silane (40 μl.) and stirred for 30 minutes. The mixturewas quenched with a 0.4M hydrochloric acid solution ofmethanol:dioxane(9:1; 9.7 ml.) and stirred for 5 minutes. The volatileswere removed in vacuo (Rotovap) and the residue was partitioned betweenwater and ethyl acetate. The separated ethyl acetate layer was washedwith water and the combined aqueous phase washed with ethyl acetate. Theaqueous phase was cooled to 0° C. and the pH adjusted to 10.3 (monitoredwith pH meter) with 1.0N sodium hydroxide. The aqueous phase wasextracted with methylene chloride (three times) and the aqueous phasewas then saturated with salt and extracted again with methylene chloride(three times). The pooled extracts were dried over sodium sulfate,filtered, concentrated, and dried in vacuo to yield 455 mg. of the freeamine as a clear oil. TLC (10% methanol in methylene chloride) R_(f)=0.28.

This free amine was dissolved in ethyl acetate (5 ml.) and treated witha solution of p-toluenesulfonic acid monohydrate (354 mg., 1 eq.) inethyl acetate (1 ml.). White crystals immediately formed. The cystalswere stored in refrigerator (5° C.) for 30 minutes and then collected byfiltration washing well with ethyl ether and drying overnight in vacuoto yield 639 mg. of title product as a white solid.

b) (S)-2-(Acetylthio)-3-cyclohexylpropanoic acid, dicyclhexylamine salt

A solution of D-phenylalanine (5.20 g., 31.5 mmol.) in 2M hydrochloricacid (75 ml.) in a 500 ml. Parr hydrogenation flask was purged withnitrogen gas and treated with platinum oxide (640 mg., 2.82 mmol.).Hydrogenation was commenced at P_(o) =42.4 psi in the sealed flask,refilling as necessary. Total hydrogen uptake was 83.4 psi (theory 83.8psi) over 6 hours. The reaction was purged with nitrogen gas andfiltered through Celite®, washing the filter cake with hot water. Thefiltrate was concentrated to about 40 ml. and stored at 5° C. overnight.The resulting solids were collected, washed with a small amount of coldwater and dried in vacuo at 60° C. to give 5.46 g. of(R)-2-amino-3-cyclohexylpropanoic acid, hydrochloride salt.

To a stirred solution of this hydrochloride salt (2.81 g., 13.5 mmol.)in 2.5N sulfuric acid (32 ml.) at room temperature was added potassiumbromide (10.0 g., 84 mmol.). The reaction mixture was cooled to -4° C.and solid sodium nitrite (1.75 g., 25.4 mmol.) was added portionwiseover one hour, maintaining the temperature below 0° C. The reactionfoamed and an oil began to form. After addition was complete, thereaction was stirred for 1 hour and then warmed to room temperature andstirred for another hour. The reaction mixture was then extracted twicewith ether, the extracts were dried (magnesium sulfate), filtered andevaporated to give 2.3 g. of (R)-2-bromo-3-cyclohexylpropanoic acid as acolorless oil.

To a stirred slurry of potassium thioacetate (1.07 g., 9.36 mmol.) indry acetonitrile (15 ml.) at 0° C. under argon was added a solution of(R)-2-bromo-3-cyclohexylpropanoic acid (2.20 g, 9.36 mmol) inacetonitrile (3 ml.) over 10 minutes. The reaction was warmed to roomtemperature and stirred 16 hours. The resulting slurry was filtered andevaporated. The residue was redissolved in ethyl acetate, washed oncewith 5% potassium bisulfate solution, dried (sodium sulfate) andevaporated. The oily yellow residue (2.21 g.) was dissolved in ether andtreated with a solution of dicyclohexylamine (1.8 ml., 9.0 mmol.) in 5ml. of ether. Scratching the flask surface with a Glass rod provided2.38 g. of white crystalline title product; m.p. 159°-161° C., [α]_(D)=-41.2° (c=1.0, chloroform).

Anal. calc'd. for C₂₃ H₄₁ NSO₃ : C, 67.11; H, 10.04; N, 3.40; S, 7.79Found: C, 66.95; H, 10.12; N, 3.25; S, 7.89.

[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-3-cylohexyl-1-oxopyrrolo[amino]-5-oxopyrrolo[2,1-b][1,3-thiazepine-7-carboxylicacid, methyl ester

A suspension of the compound (S)-2-(acetylthio)-3-cyclohexylpropanoicacid, dicyclohexyamine salt (285 mg., 0.69 mol., 1.05 eq.) in ethylacetate (15 ml.) was washed with 5% potassium bisulfate (3×10 ml.), 50%brine (10 ml.), and brine (10 ml.), dried (anhydrous magnesium sulfate),filtered, concentrated, stripped with methylene chloride (twice) anddried in vacuo for one hour to give(S)-2-(acetylthio)-3-cyclohexylpropanoic acid as an oil.

This free acid of was dissolved in dry methylene chloride (5 ml.),cooled to 0° C. (ice bath), and treated with triethylamine (96 μl., 0.69mmol., 1.05 eq.), then the product from part (a) (275 mg., 0.66 mmol.),triethylamine (92 μl., 0.66 mmol.) and finallybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(305 mg., 0.69 mmol.). The resultant solution was stirred at 0° C. for 1hour then at room temperature for 2 hours. The reaction mixture wasconcentrated, diluted with ethyl acetate, washed with 5% potassiumbisulfate (20 ml.), 50% saturated sodium bicarbonate (20 ml.), brine (20ml.), dried (anhydrous magnesium sulfate), filtered, and evaporated todryness. The crude product was adsorbed onto Celite® and chromatographedon a silica gel column (2.5×10 cm), eluting with 40% (1 l.) ethylacetate/hexane. The desired fractions were combined and concentrated,affording 246 mg. of pure title product. TLC (ethyl acetate:hexane, 8:2)R_(f) =0.53.

d)[4S-[4α(R*),7α9aβ]]-Octahydro-4-[(3-Cyclohexyl-2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid

A solution of the product from part (c) (245 mg., 0.54 mmol.) inmethanol (6 ml.), purged with argon for 30 minutes, cooled to 0° C. wastreated dropwise with a previously purged (argon, 30 minutes) solutionof 1.0N sodium hydroxide (5 ml.) maintaining the bubbling of argonthroughout the addition and length of the reaction. The reaction mixturewas stirred at 0° C. for 2 hours, acidified at 0° C. with 5% potassiumbisulfate to pH 2 then extracted with ethyl acetate (3×20 ml.). Thecombined organic extracts were washed with 50% brine (20 ml.), and brine(20 ml.), dried (anhydrous magnesium sulfate), filtered, and evaporatedto dryness. The residue was purified by chromatography on a 2.5×10 cmsilica gel column eluting with 7:3 ethyl acetate:heptane (300 ml.) and1% acetic acid in 7:3 ethyl acetate:heptane (500 ml.). The desiredfractions were concentrated, stripped with methylene chloride and driedin vacuo to yield 172 mg. of title product as a white foam; [α]_(D)=-116.9° (c=0.5, methanol). TLC (1% acetic acid in ethyl acetate) R_(f)=0.35.

¹ H-NMR: 400 MHz; CDCl₃ : δ0.91 (m, 2H), 1.22 (m, 3H), 1.44 (m, 1H),1.55 (m, 1H), 1.68 (m's, 5H), 1.83 (m, 1H), 1.97 (m's, 2H), 2.12 (m,1H), 2.19-2.40 (m's, 3H), 2.53 (m, 1H), 2.96 (m, 1H), 3.38 (m's, 2H),4.62 (t, 1H, J=6.8 Hz), 4.70 (m, 1H), 5.25 (m, 1H), 7.55 (d, 1H, J=6.4Hz).

¹³ C-NMR: 100 MHz; CDCl₃ : δ26.0, 26.1, 27.5, 31.5, 32.3, 33.0, 33.3,35.2, 40.7, 43.0, 52.9, 60.6, 62.5,170.9, 172.7, 175.3.

Anal. calc'd. for C₁₈ H₂₈ N₂ O₄ S₂ : C, 53.98; H, 7.05; N, 6.99; S,16.01 Found: C, 53.97; H, 7.18; N, 6.84, S, 15.75. HPLC: t_(R) =16 min(>99%, UV 217); YMC S-3 ODS (C-18) 6.0×150 mm; 50% B:A-100% B:A, 25minute linear gradient (A=90% water/methanol+0.2% phosphoric acid; B=90%methanol/water+0.2% phosphoric acid); flow rate at 1.5 ml/min.

EXAMPLE 8[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxohexyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

a) (S)-2-Bromohexanoic acid

Potassium bromide (15.9 g., 133 mmol.) was added to a stirred solutionof D-norleucine (5.0 g., 38 mmol.) in 2.5N sulfuric acid (77 ml.) atroom temperature. The reaction mixture was cooled to -10° C. and solidsodium nitrite (3.94 g., 57 mmol.) was added portionwise, maintainingthe temperature between -10° and -5° C. After addition was complete, thefoamy reaction was stirred for 1 hour and then warmed to roomtemperature and stirred for another hour. The reaction mixture was thenextracted twice with ether, the ether extracts were washed once withwater, dried (magnesium sulfate), filtered and evaporated to give 3.3 g.of crude title product.

b) (S)-2-(Acetylthio)hexanoic acid, dicyclohexylamine salt

To a stirred slurry of potassium thioacetate (2.11 g., 18.5 mmol.) in 50ml. of dry acetonitrile at room temperature under argon was added asolution of the product from part (a) (3.27 g., 16.8 mmol.) in 26 ml. ofacetonitrile. The reaction was stirred 5 hours. The resulting slurry wasfiltered and evaporated. The residue was redissolved in ethyl ether,washed once with 5% potassium bisulfate solution and once with brine,dried (magnesium sulfate) and evaporated. The residue was dissolved inether (64 ml.) and treated with dicyclohexylamine 3.4 ml., 16.8 mmol.).The ethereal solution was concentrated in vacuo, and triturated fromhexanes to give a white solid which was recrystallized from ethylether/hexanes to give the title product. The mother liquor wasconcentrated and recrystallized twice to provide a total yield of 2.2 g.of title product; m.p. 145°-147° C.; [α]_(D) =-33.8° (c=1.08,chloroform).

c)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxohexyl]amino]-5-oxopyrrolo-[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A suspension of the dicyclohexylamine salt product from part (b) (255mg., 0.69 mmol., 1.05 eq.) in ethyl acetate (15 ml.) was washed with 5%potassium bisulfate (3×5 ml.), and brine (10 ml.), dried (anhydrousmagnesium sulfate), filtered, concentrated, stripped with methylenechloride (twice), and dried in vacuo for one hour to give the free acidas a oil.

This oil was dissolved in dry methylene chloride (6 ml.), cooled to 0°C. (ice bath) and treated with triethylamine (96 μl., 0.69 mmol., 1.05eq.), then[4S-(4α,7α,9aβ)]-4-amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [prepared as describedin Example 7(a), 275 mg., 0.66 mmol.], triethylamine (92 μl., 0.66mmol.) and finally benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (305 mg., 0.69 mmol.). The resultant solution wasstirred at 0° C. for one hour then at room temperature for 2 hours. Thereaction mixture was concentrated, diluted with ethyl acetate, washedwith 5% potassium bisulfate (20 ml.), 50% saturated sodium bicarbonate(20 ml.), brine (20 ml.), dried (anhydrous magnesium sulfate), filtered,and evaporated to dryness. The crude product was adsorbed onto Celite®and chromatographed on a silica gel column (2.5×10 cm), eluting with 40%(1 l.) ethyl acetate/hexane. The desired fractions were combined andconcentrated, affording 258 mg of pure title product. TLC (ethylacetate:hexane, 8:2) R_(f) =0.54.

d)4S-[4α(R*),7α,9aβ]]-Ocxtahydro-4-[2-mercapto-1-oxohexyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (c) (255 mg., 0.54 mmol.) inmethanol (5 ml.), purged with argon for 30 minutes, cooled to 0° C. wastreated dropwise with a previously purged (argon, 30 minutes) solutionof 1.0N sodium hydroxide (5 ml.) maintaining the bubbling of argonthroughout the addition and length of the reaction. The reaction mixturewas stirred at 0° C. for 2 hours, acidified at 0° C. with 5% potassiumbisulfate to pH 2 then extracted with ethyl acetate (3×20 ml.). Thecombined organic extracts were washed with 50% brine (20 ml.) and brine(20 ml.), dried (anhydrous magnesium sulfate), filtered, and evaporatedto dryness. The residue was purified by chromatography on a 2.5×10 cmsilica gel column eluting with 7:3 ethyl acetate:heptane (300 ml.) and1% acetic acid in 7:3 ethyl acetate:heptane (500 ml.). The desiredfractions were concentrated, stripped with methylene chloride and driedin vacuo to yield 170 mg. of title product as a white foam; [α]_(D)=-135.1° (c=0.5, methanol). TLC (1% acetic acid in ethyl acetate) R_(f)=0.32.

¹ H-NMR: 400 MHz; CDCl₃ : δ0.89(t, 3H, J=7 Hz), 1.32(m, 4H), 1.73(m,1H), 1.96(m, 2H), 2.00(d, 1H, J=8.6 Hz ), 2.11(m, 1H), 2.32 (m's, 3H),2.52(m, 1H), 2.98(m, 1H), 3.32(m's, 2H), 4.61 (t, 1H, J=7.1 Hz), 4.72(m, 1H), 5.25 (m, 1H), 7.63 (d, 1H, J=6.4 Hz).

¹³ C-NMR: 100 MHz; CDCl₃ : δ13.8, 22.2, 27.6, 29.2, 31.4, 32.6, 33.1,35.3, 43.0, 52.8, 60.5, 62.42, 170.7, 172.5. 174.0.

Anal. calc'd for C₁₅ H₂₄ O₄ N₄ S₂ O•0.08 H₂ O: C, 49.79; H, 6.73; N,7.74; S, 17.72 Found: C, 49.90; H, 6.92; N, 7.63, S, 17.57. HPLC: t_(R)=9.4 min (>99%, UV 220); YMC S-3ODS (C-18) 6.0×150 mm; 50% B:A-100% B:A,25 minute linear gradient (A=90% water/methanol+0.2% phosphoric acid);B=90% methanol/water+0.2% phosphoric acid); flow rate at 1.5 ml/min.

EXAMPLE 9[4S-4α(R*),7α,9aβ]]-Ocatahydro-4-[(2-mercapto-1-oxo-4-methylpentyl)amino]-5-oxopyrrolo[2,1-b][1,3-thiazepine-7-carboxylicacid

a) (S)-2-Bromo-4-methytlpentanoic acid

Potassium bromide (9.5 g., 80 mmol.) was added to a stirred solution ofD-leucine (3.0 g., 23 mmol.) in 2.5N sulfuric acid (47 ml.) at roomtemperature. The reaction mixture was cooled to -10° C. and solid sodiumnitrite (2.4 g., 34 mmol.) was added portionwise, maintaining thetemperature between -10° and -5° C. After addition was complete, thereaction was stirred for 1 hour and then warmed to room temperature andstirred for another hour. The reaction mixture was then extracted twicewith ether, the ether extracts were washed once with water, dried(magnesium sulfate), filtered and evaporated to give 2.7 g. of crudetitle product.

b) (S)-2-(Acetylthio)-4-methylpentanoic acid, dicyclohexylamine salt

To a stirred slurry, of potassium thioacetate (1.7 g., 15.0 mmol.) in 50ml. of dry acetonitrile at room temperature under argon was added asolution of the product from part (a) (2.6 g., 13 mmol.) in 17 ml. ofacetonitrile. The reaction was stirred 4 hours. The resulting slurry wasfiltered and evaporated. The residue was redissolved in ethyl ether,washed once with 5% potassium hydrogen sulfate solution and once withbrine, dried (magnesium sulfate) and evaporated. The residue wasdissolved in ether (64 ml.) and treated with dicyclohexylamine (2.7 ml.,14 mmol.). A white solid immediately began precipitating from thesolution. The solution was filtered and the white solid collected togive 2.0 g. of title product; m.p. 153°-158° C.; [α]_(D) =-54.5° C.(c=0.61, chloroform).

c)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylyhio)-1-oxo-4-methylpentyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A stirred suspension of the dicyclohexylamine salt product from part (b)(234 mg., 0.63 mmol.) in ethyl acetate (15 ml.) was washed with 5%aqueous potassium bisulfate (3×8 ml.). The organic extract was dried(anhydrous magnesium sulfate), filtered and evaporated twice fromhexane. The resulting oil was dissolved in methylene chloride (6 ml.)and stirred under nitrogen at 0° C. To this solution was addedtriethylamine (88 μl., 0.63 mmol.), then[4S-(4α,7α,9aβ)]-4-amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylic acid, methyl ester, p-toluenesulfonicacid salt [prepared as described in Example 7(a), 249 ml., 0.6 mmol.],an additional amount of triethylamine (84 μl., 0.60 mmol.) and, after 10minutes, benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (279 mg., 0.63 mmol.). After one hour, the reactionwas warmed to room temperature and stirred 2 hours. The resultingcolorless solution was evaporated at less than 30° C. and the oilyresidue redissolved in ethyl acetate. The solution was washed once with5% potassium bisulfate solution once with saturated sodium bicarbonatesolution and once with brine. The organic layer was dried (magnesiumsulfate), filtered and evaporated onto 5 g. of silica gel. Purificationby flash chromatography (2.5×15 cm column, eluting with 1:1 ethylacetate/hexanes) provided 203 mg. of title product as a white solid;m.p. 101°-103° C. [α]_(D) =-157.5° (c=1.04, chloroform). TLC (ethylacetate:hexane, 1:1) R_(f) =0.19.

d)[4S-[4α(R*),7α,9aβ]]-Ocytahydro-4-[(2-mercapto-1-oxo-4-methylpentyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (c) (184 mg., 0.44 mmol.) in 5 ml.of methanol was purged with nitrogen for 10 minutes and cooled to 0° C.To this solution was added dropwise 5 ml. of nitrogen-purged 1M sodiumhydroxide. Nitrogen was slowly bubbled through the solution during thereaction. After 2 hours, the reaction was acidified with 2 ml. of 6Mhydrochloric acid, extracted twice with ethyl acetate and the extractscombined, dried (magnesium sulfate) and evaporated. Re-evaporation fromhexanes and trituration of the residue in methanol/water provided 132mg. of title product as a crystalline solid, m.p. 94°-96° C.; [α]_(D)=-158.6° (c=0.42, methanol). TLC(ethyl acetate:hexane:acetic acid,4:4:0.1) R_(f) =0.13.

Anal. calc'd. for C₁₅ H₂₄ N₂ S₂ O₄ •0.75 H₂ O: C, 48.17; H, 6.87; N,7.49; S, 17.15 Found: C, 48.33; H, 6.51; N, 7.37; S, 16.82. HPLC: R_(t)=17.6 min; (99.2%) YMC S-3 ODS (C-18) 6.0×150 mm; 0% to 100% B:A, 25 minlinear gradient and 15 min hold, 1.5 mL/min; A=90% water/methanol+0.2%phosphoric acid; B=90% methanol/water+0.2% phosphoric acid; 220 nm.

EXAMPLE 10[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo[1,4]oxozina[3,4-b][1,3]-oxazepine-7-carboxylicacid

a) 2,2,2-Trichloroacetimidoic acid, 2-propenyl ester

A suspension of 80% sodium hydride (945 mg., 31.5 mmol.; washed twicewith 25 ml. of hexane) in dry. ether (30 ml.) was treated dropwise witha solution of 2-propen-1-ol (21.4 ml., 18.3 g., 315 mmol.), in dry ether(45 ml.), stirred for 20 minutes at room temperature under argon andthen cooled to 0° C. (ice-salt bath). Trichloroacetonitrile (30 ml. or42.3 g., 0.30 mole) was added over a period of 15 minutes and thebrownish solution was stirred at 0° C. for 40 minutes, at 10° C. for 10minutes and at room temperature for 10 minutes. The reaction mixture wasconcentrated to a syrup, treated with a solution of methanol (1.2 ml.)in pentane (30 ml.) and stirred vigorously for 5.0 minutes. The lightbrown precipitates were filtered off, washed with pentane (2×30 ml.) andthe combined flitrates concentrated down to a light brown liquid. Theliquid was redissolved in pentane (30 ml.), stirred for a few minutes,and the resulting suspension filtered, and the precipitates obtainedwashed with pentane (30 ml.), repeating the procedure at least one moretime. The clear filtrate was concentrated and dried in vacuo to give54.0 g. of title compound as a light red-colored liquid. This materialwas stored as a solution in hexane at 10° C.

b) N-Phthaloyl-L-serine, methyl ester

A suspension of L-serine, methyl ester, hydrochloride, (25 g., 161mmol.) in water (350 ml.) was diluted with dioxane (250 ml.) and theresulting clear solution treated with solid sodium carbonate (17 g., 1.0eq.) followed by N-carbethoxyphthalimide (37 g., 1.05 eq.). The reactionmixture was stirred at room temperature for 2.5 hours under argon. Themixture was extracted with ethyl acetate (3×500 ml.) and the combinedorganic extracts were washed successively with 5% sodium bicarbonate(250 ml.), 5% potassium bisulfate (250 ml.) and brine (250 ml.), dried(anhydrous sodium sulfate), filtered, evaporated to dryness and dried invacuo. The product mixture was chromatographed on a silica gel column(Merck), eluting the column with ethyl acetate:hexane mixtures (1:3;1:2) and the desired fractions were combined, evaporated to dryness anddried in vacuo to give 31 g. of title compound as a thick syrup. TLC(ethyl acetate:hexane, 1:1) R_(f) =0.52.

c) N-Phthaloyl-O-(2-propenyl)-L-serine, methyl ester

A solution of the product from part (b) (7.37 g., 29.5 mmol.) in drymethylene chloride (30 ml.) was treated with a solution of the productfrom part (a) (11.97 g., 59.1 mmol., 2 eq.) in cyclohexane (60 ml.)followed by trifluoromethanesulonic acid (0.37 ml.) and the reactionmixture was stirred at room temperature for 20 hours under argon. Theprecipitates were filtered off, washed with a minimal amount ofmethylene chloride and the combined flitrates were washed with 5% sodiumbicarbonate (30 ml.) and water (30 ml.), dried (anhydrous sodiumsulfate), filtered, evaporated to dryness and dried in vacuo. The crudeproduct mixture was chromatographed on a silica gel column (Merck),eluting the column with ethyl acetate:hexane (1:9). The desiredfractions were combined, evaporated to dryness and dried in vacuo togive 7.56 g. of title compound as a clear thick syrup. TLC(ethylacetate: hexane, 1:1) R_(f) =0.70.

d) N-Phthaloyl-O-(acetaldehyde)-L-serine, methyl ester

A solution of the product from part (c) (2.5 g., 8.64 mmol.) in amixture of dry methylene chloride (46.4 ml.) and methanol (4.6 ml.) wascooled to -78° C. (dry ice-acetone bath) and treated with ozone until ablue color persisted (about 15 minutes). The mixture was then purgedwith nitrogen for 10 minutes (until the blue color disappeared), treatedwith dimethylsulfide (14.0 ml., 0.19 mole, 22.1 eq.), warmed to roomtemperature and stirred for 2.5 hours under nitrogen. The reactionmixture was evaporated to dryness and the residual syrup dissolved inethyl acetate (50 ml.), washed with water (15 ml.) and brine (15 ml.),dried (anhydrous magnesium sulfate), filtered, evaporated to dryness anddried in vacuo.

The crude product was chromatographed on a silica gel column (Merck),eluting the column with ethyl acetate: hexane mixtures (1:9; 1:4; 1:2 )to give 1.54 g. of title product. TLC(ethyl acetate:hexane, 1:1) R_(f)=0.33.

e) N-Phthaloyl-O-(2,2-dimethoxyethyl)-L-serine, methyl ester

A solution of the product from part (d) (1.54 g., 5.29 mmol.) in amixture of dry methylene chloride (8.3 ml.) and dry methanol (8.3 ml.),was treated with trimethylorthoformate (0.84 ml., 7.68 mmol., 1.45 eq.)and p-toluenesulfonic acid monohydrate (92 mg.). The reaction mixturewas stirred at room temperature under argon for 2.5 hours thenpartitioned between ethyl acetate (50 ml.) and saturated sodiumbicarbonate (15 ml.). The organic phase was washed with water (15 ml.)and brine (15 ml.), dried (anhydrous sodium sulfate), filtered,evaporated to dryness and dried in vacuo. The crude product waschromatographed on a silica gel column (Merck), eluting the column withethyl acetate:hexane (1:4) to give 1.35 g. of title product as a thickclear syrup. TLC(ethyl acetate:hexane, 1:1) R_(f) =0.58.

f) O-(2,2-Dimethoxyethyl)-L-serine, methyl ester

A solution of the product from part (e) (2.0 g., 5.93 mmol.) in drymethanol (14 ml.) was treated with hydrazine hydrate (0.30 ml., 6.1mmol.) and stirred at room temperature for 4 days under argon. Theresulting suspension was filtered, washing the precipitates withmethanol (2×14 ml.) and the filtrate was concentrated to dryness. Thesyrup was re-dissolved in methylene chloride and filtered two more timesuntil no more precipitates were obtained. The clear filtrate wasconcentrated to give 1.17 g. of title product as a light yellow syrup.TLC(methylene chloride:methanol, 9:1) R_(f) =0.54.

g) N-[(Phenylmethoxy) carbonyl]-O-[(1,1-dimethylethyl)dimethylsilyl]-L-homoserine

A solution of N-[(phenylmethoxy)carbonyl]-L-homoserine [prepared asdescribed in Example 6(a), 3.0 g., 11.85 mmol.] in dry dimethylformamide(65 ml.) was treated with [(1,1-dimethylethyl)dimethylsilyl chloride(10.72 g., 71.1 mmol.) and imidazole (9.65 g, 0.14 mol.) and stirred atroom temperature under argon for 24 hours. The reaction mixture wasdiluted with methanol (207 ml.), stirred for another 24 hours at roomtemperature and then concentrated to a syrup. The residual syrup wasdissolved in ethyl acetate (200 ml.), washed with 10% citric acid (2×75ml) and brine, dried (anhydrous sodium sulfate), filtered, evaporated todryness and dried in vacuo. The crude product mixture waschromatographed on a silica gel column (Merck), eluting the column withethyl acetate:hexane (1:1) followed by ethyl acetate:acetic acid(99.5:0.5). The desired fractions were combined, concentrated andevaporated several times from toluene to give 3.56 g. of title compoundas a waxy solid. TLC(ethyl acetate:acetic acid, 95:5) R_(f) =0.82.

h)N-[O-(1,1-Dimethylethyl)dimethylsilyl]-N-[(phenylmethoxy)carbonyl]-L-homoseryl]-O-(2,2-dimethoxyethyl)-L-serine,methyl ester

A solution of the product from part (g) (2.18 g., 5.93 mmol.) in drymethylene chloride was cooled to 0° C. (ice-salt bath) and treatedsequentially with a solution of the product from part (f) (1.71 g., 5.65mmol.) in dry methylene chloride (5 ml), triethylamine (0.78 ml., 5.65mmol.) and benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (2.63 g., 6.0 mmol.). The reaction mixture wasstirred at 0° C. for 30 minutes and at room temperature for one hour and45 minutes. The reaction mixture was partitioned between ethyl ether(2×100 ml.) and water (30 ml.) and the combined organic extracts werewashed with 50% saturated sodium bicarbonate (20 ml.) and brine (25ml.), dried (anhydrous sodium sulfate), filtered, evaporated to drynessand dried in vacuo. The crude product mixture was chromatographed on asilica gel column (Merck), eluting the column with ethyl acetate:hexanemixtures (1:4; 1:3; 1:1) to give 2.38 g. of title product. TLC(ethylacetate:hexane, 1:1) R_(f) =0.40.

i)[4S-(4α,7α,10aβ)]-Octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-oxo[1,4]oxazino[3,4-b][1,3]-oxazepine-7-carboxylicacid, methyl ester

A solution of the product from part (h) (1.0 g., 1.8 mmol.) in drymethylene chloride (50 ml.) was treated with Amberlyst® 15 ion exchangeresin (acid form) and methanol (0.1 ml.) and the resulting mixture wasstirred at room temperature under argon for three days. The resin wasfiltered off, washed with a small amount of methylene chloride and thefiltrate concentrated to a syrup. The crude product mixture waschromatographed on a silica gel column, eluting the column with ethylacetate:hexane (1:1) to give 339 mg. of title product. TLC(ethylacetate:hexane, 3:1) R_(f) =0.53.

j)[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo[1,4]oxazino[3,4-b][1,3]oxazepine-7-carboxylicacid, methyl ester

A solution of the product prepared as described part (i) (771 mg., 2.04mmol.) in dry methanol (25 ml.) was treated with 10% palladium on carboncatalyst (125 mg.) and hydrogenated (balloon) at room temperature for 16hours. The reaction mixture was filtered through a Celite® pad and thepad was washed with methanol (2×25 ml.). The clear filtrate wasevaporated to dryness and dried in vacuo to give 448 mg. of titleproduct as a syrup. TLC(methylene chloride:methanol, 9:1) R_(f) =0.22.

k)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetythio)-1-oxo-3-phenylpropyl]amino]5-oxo[1,4]oxazino[3,4-b][1,3]oxazepine-7-carboxylicacid, methyl ester

The dicyclohexylamine salt of (S)-2-(acetylthio)benzenepropanoic acid(813 mg., 2.01 mmol.) was suspended in ethyl acetate (70 ml.), washedwith 5% potassium bisulfate (5×9.3 ml.) and brine (9.3 ml.), dried(anhydrous magnesium sulfate), filtered, evaporated to dryness and driedin vacuo.

This free acid was dissolved in dry methylene chloride (12 ml.), cooledto 0° C. (ice-salt bath) and treated sequentially with a solution of theproduct from part (j) (448 mg., 1.84 mmol.) in dry methylene chloride(4.0 ml.), triethylamine (0.25 ml., 1.80 mmol.) andbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(823 mg., 1.86 mmol.). The reaction mixture was stirred at 0° C. for onehour and at room temperature for 2 hours under argon. The reactionmixture was stripped to dryness and the syrup obtained was re-dissolvedin ethyl acetate (60 ml.), washed with 0.5N hydrochloric acid (2×11ml.), water (11 ml.) and brine (11 ml.), dried (anhydrous sodiumsulfate), filtered, evaporated to dryness and dried in vacuo. The crudeproduct mixture was chromatographed twice on a silica gel column(Merck), eluting each column with ethyl acetate:hexane mixtures (1:1;1:2) to give 665 mg. of title product as a syrup. TLC(ethylacetate:hexane, 3:1) R_(f) =0.30.

l)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo[1,4]oxazino[3,4-b]-[1,3]oxazepine-7-carboxylicacid

A solution of the product from part (k) (650 mg., 1.44 mmol.) inmethanol (13 ml.), was purged with argon for 30 minutes, cooled down to0° C. (ice-salt bath) and treated dropwise with a solution of 1.0Nsodium hydroxide (5.84 ml., previously purged with argon for 30minutes), maintaining the bubbling of argon throughout the addition andlength of the reaction. The reaction mixture was stirred at 0° C. for5.0 hours and quenched at 0° C. with 5% potassium bisulfate (25.4 ml.).The mixture was warmed to room temperature, extracted with ethyl acetate(3×50 ml.) and the combined organic extracts were washed with brine (15ml.), dried (anhydrous sodium sulfate), filtered, evaporated to drynessand dried in vacuo. The crude product was triturated withhexane:methylene chloride (130:7) and the solid obtained chromatographedon a silica gel column (Merck), eluting the column with ethylacetate:hexane mixtures (1:2; 1:1) followed by methylenechloride:methanol:acetic acid (100:4:0.2). The desired fractions werecombined, evaporated to dryness and evaporated several times fromtoluene to give 364 mg. of title product which was dried in vacuo for9.0 hours. The resulting product was then triturated with methylenechloride:hexane (1:10), hexane (50 ml.) and pentane (2×50 ml.), stirringwith the first 50 ml. for 4 hours and the next 50 ml. of pentaneovernight under argon. The solvent was decanted and the solid dried invacuo for 6.0 hours to give pure title product as a solid amorphousfoam; [α]_(D) =-49.1° (c=0.48, methanol). TLC (toluene:acetic acid, 5:1)R_(f) =0.17.

Anal. calc'd. for C₁₈ H₂₂ N₂ O₆ S•0.56 H₂ O: C, 53.45; H, 5.76; N, 6.93;S, 7.92 Found: C, 53.45; H, 5.53; N, 6.75; S, 7.48. HPLC: R_(t) =10.45min.; (98.3%); YMS S-3 ODS (c=18) 6.0×150 mm; 44% (10% water-90%methanol-0.2% phosphoric acid)/56% (90% water-10% methanol-0.2%phosphoric acid), isocratic; 1.5 ml/min.

EXAMPLE 11[4S-[4α(R*),7α,10aβ]]-Octahydro-4-(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrrolo[2,1-b][1,3]-thiazepine-7-carboxylicacid

The product of Example 3 was also prepared as follows:

a)N-[(Phenylmethoxy)carbonyl]-O-[(1,1-dimethylethyl)dimethylsilyl]-L-homoserine

[(1,1-Dimethylethyl)dimethylsilyl]chloride (37.5 g., 249 mmol.) wasadded to a solution of N-[(phenylmethoxy)carbonyl]-L-homoserine[prepared as described in Example 6(a), 41.56 mmol.] indimethylformamide (125 ml.), followed by imidazole (33.95 g., 498mmol.). The resulting light yellow solution was stirred at roomtemperature for 22 hours. Methanol (500 ml.) was added, the reactionmixture was stirred for an additional 6 hours, and then the methanol andmost of the dimethylformamide were removed in vacuo. The remainingresidue was taken up into ethyl acetate (800 ml.), washed with 10%citric acid (2×300 ml.), and the combined aqueous phase was extractedwith ethyl acetate (300 ml.). The combined ethyl acetate phase waswashed with water and brine, dried (sodium sulfate), concentrated, andthe residue was evaporated with hexane to form a white powder. Thispowder was dried in vacuo to give 12.942 g. of title product.

b)[S-(R*,R*)]-2-[[-4-[[(1,1-Dimethylethyl)dimethylsilyl]oxy]-1-oxo]-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

To a solution of the product from part (a) (22.78 g., 61.97 mmol.) inmethylene chloride (100 ml.) cooled at 0° C. was addedN-methylmorpholine (6.81 ml., 61.97 mmol.), followed byhydroxybenzotriazole (8.37 g., 61.97 mmol.),(S)-2-amino-6,6-dimethoxyhexanoic acid, methyl ester [prepared asdescribed in Example 1(e), 10.6 g., 51.64 mmol.] in methylene chloride(50 ml.), and then 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (11.88g., 61.97 mmol.). The reaction mixture was stirred at 0° C. for onehour, then at room temperature overnight. The reaction mixture wasconcentrated in vacuo and the residue was diluted with ethyl acetate(600 ml.), washed with 5% potassium bisulfate (200 ml.), 0.5N sodiumhydroxide (200 ml.), water, and brine, and dried (sodium sulfate). Thefiltrate was concentrated and the residue taken up in ethyl ether (150ml.). The resulting suspension was filtered and the collected solid waswashed thoroughly with ethyl ether. The filtrate was concentrated invacuo to dryness to afford 30 g. of crude title product as an oilycompound which was used in the next reaction without purification. TLC(8:2, ethyl acetate:hexane) R_(f) =0.55.

c)[S-(R*,R*)]-2-[[4-Hydroxy-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

To a solution of the product from part (b) (30 g.) in methanol (150 ml,)cooled at 0° C. was added para-toluenesulfonic acid monohydrate (1.96g,), The reaction mixture was stirred at 0° C. for 2 hours beforequenching with aqueous sodium bicarbonate solution (1.3 g. of sodiumbicarbonate in 100 ml. of water). The mixture was concentrated in vacuoand the residue partitioned between ethyl acetate (400 ml.) and water(150 ml.). The separated aqueous phase was extracted with ethyl acetate(2×150 ml.). The combined ethyl acetate layers were washed with 10%sodium bicarbonate, brine (2 times), dried (sodium sulfate), filteredand evaporated to dryness. The residue was flash chromatographed on a10×25 cm silica gel column eluting with 80% ethyl acetate in hexane(51.), ethyl acetate (31.) and 2% methanol in ethyl acetate (51.). Thedesired fractions were combined and concentrated, and dried in vacuo togive 17.45 g. of title product as a pale yellow oil. TLC (8:2, ethylacetate:hexane) R_(f) =0.17.

d)[S-(R*,R*)]-2-[[-4-[(Methanesulfonyl)oxy]-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

To a solution of the product from part (c) (17.40 g., 39.50 mmol.)(stripped with toluene three times and dried in vacuo overnight) in drymethylene chloride (250 ml.) cooled at -15° C. (ice/acetone) was addedtriethylamine (8.26 ml., 59.28 mmol, freshly distilled), followed bymethanesulfonyl chloride (3.67 ml., 47.4 mmol.) dropwise. The reactionmixture was stirred at -15° C. for 30 minutes, then quenched withsaturated ammonium chloride solution (100 ml.). After stirring for 5minutes, the mixture was diluted with ethyl acetate (600 ml.) and washedwith 5% potassium bisulfate, brine, dried (sodium sulfate), filtered andevaporated to dryness. The residue was dried in vacuo to give 20.40 g.of title compound as a yellow oil which was used in the next reactionwithout purification. TLC (8:2, ethyl acetate: hexane) R_(f) =0.35.

e)[S-(R*,R*)]-2-[[4-(Acetythio)-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

To a solution of thioacetic acid (5.09 ml., 71.10 mmol.) in methanol(100 ml.) was added cesium carbonate (10.81 g., 33.18 mmol.). Theresulting solution was concentrated in vacuo. The solid was trituratedwith dry acetone (3 times) and then dried in vacuo over phosphoruspentoxide overnight to give cesium thioacetate.

A solution of the product from part (d) (20.40 g., 39.50 mmol.) in dry.dimethylformamide (150 ml.) was added via cannula to a suspension ofcesium thioacetate (10.576 g., 50.85 mmol.) in dimethylformamide (50ml.). The resulting yellow solution was stirred under argon at roomtemperature overnight, then concentrated at high vacuum to remove mostof the dimethyl formamide. The residue was taken into ethyl acetate(11.) and washed with 10% sodium bicarbonate (200 ml.), water (4×200ml.), brine (400 ml.) and dried (sodium sulfate). The filtrate wasconcentrated and the residue evaporated with toluene (3 times), thendried in vacuo to afford 20 g. of title compound as a light yellow solidwhich was used for the next reaction without purification. TLC(8:2;ethyl acetate:hexane) R_(f) =0.47.

f)[S-(R*,R*)]-2-[[4-Mercapto-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

A solution of the product from part (e) (20 g., 39.50 mmol.) in methanol(250 ml.) cooled at 0° C. was purged with argon for 15 minutes. Withcontinuous argon purging, a 25% (weight/weight, density=0.945) sodiummethoxide solution in methanol (9.17 ml., 40 mmol.) was added dropwise.After stirring for 5 minutes, the reaction was quenched with saturatedammonium chloride solution (200 ml.) and the mixture partitioned betweenethyl acetate (11.) and water (200 ml.). The aqueous phase was extractedwith ethyl acetate (200 ml.). The combined ethyl acetate extract waswashed with saturated ammonium chloride solution (400 ml.), brine (400ml.), dried (sodium sulfate), filtered and concentrated in. vacuo togive 17.5 g. of title product as a yellow oil which was used for thenext reaction without purification. TLC (8:2, ethyl acetate:hexane)R_(f) =0.45.

g)[4S-(4α,7α,10aβ)]-Octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

To a solution of the product from part (f) (17.5 g., 38.3 mmol.) inmethylene chloride (600 ml.) was added Amberlyst® 15 ion exchange resin(6 g., pretreated with 6N hydrochloric acid, water, tetrahydrofuran, andmethylene chloride and dried). The suspension was stirred under argon atroom temperature for 18 hours and filtered. The filtrate wasconcentrated and the residue adsorbed on Celite®, purified on a 10×30 cmsilica gel column eluting with 20-30% of ethyl acetate in hexane. Thedesired fractions were combined and evaporated in vacuo to dryness toafford 9.18 g. of title product as a yellow oil. TLC (1:1, ethylacetate:hexane) R_(f) =0.32.

h)[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

To a solution of solution of the product from part (g) (9.1 g., 23.19mmol., evaporated with toluene three times and dried in vacuo overnight)in dry methylene chloride (150 ml.) was added iodotrimethylsilane (4.95ml., 34.78 mmol.) dropwise. The resulting yellow solution was stirredunder argon at room temperature for 1.5 hours, then quenched with 0.4Nhydrochloric acid in methanol/dioxane (120 ml.). The volatiles wereremoved in vacuo and the residue partitioned between ether (500 ml.) andwater (700 ml.). The organic phase was extracted with 0.1N hydrochloricacid (150 ml.) and the combined acidic aqueous extract cooled to 0° C.,basified with 1N sodium hydroxide to pH 10.5 (monitored with a pHmeter), then extracted with methylene chloride (4×400 ml.). The combinedorganic extracts were washed with brine, dried (sodium sulfate),filtered and concentrated in vacuo to afford 6.45 g. of title product asa yellow oil, which was used for the next reaction without furtherpurification. TLC (1:9, methanol:methylene chloride) R_(f) =0.20.

i)[4S-[4α(R*),7α,10aβ[[-Octahydro-4-[2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

A cold (0° C.) solution of (S)-(2-acetylthio)benzenepropanoic acid andtriethylamine in methylene chloride was treated with a solution of theproduct from part (h) in methylene chloride followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate.The reaction was worked up according to the procedure described inExample 3(c) to afford[4S-[4α(R*),7α,10aβ]]-octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester.

A solution of this methyl ester product in deoxygenated methanol wastreated with 1N sodium hydroxide according to the procedure of Example3(d) to afford the title product.

EXAMPLE 12

a)4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-mercaptomethyl)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylicacid

a) (S)-2-[(Acetylthio)methyl]benzepropanoic acid, ephedrine salt

A solution of (1R,2S)-(-)-ephedrine (17.3 g., 105 mmol.) in diethylether (175 ml.) was added in one portion to a solution of2-[(acetylthio)methyl]benzenepropanoic acid (50.0 g., 210 mmol.) indiethyl ether (175 ml.). After standing at room temperature for 16hours, the crystallized ephedrine salt was collected by filtration (19.7g.); m.p. 114°-125°; [α]_(D) =-40.6° (c=1, methanol). An additionalamount of solid [8.9 g, m.p. 121°-126°; [α]_(D) =-47.2° (c=1, methanol)]separated from the filtrate after remaining at room temperature for 20hours. The solids were combined and recrystallized from acetonitrile(1500 ml.). After 16 hours at room temperature, 20.8 g. of solid wascollected; m.p. 125°-130° C.; [α]_(D) =-48.9° (c=1, methanol). Thismaterial was recrystallized in the same manner from acetonitrile (300ml.) to give 18.7 g., m.p. 128°-130° ; [α]_(D) =-48.9° (c=1, methanol).A third recrystallization from acetonitrile (225 ml.) afforded 17.4 g ofsolid (S)-2-[(acetylthio)methyl]benzenepropanoic acid, ephedrine salt;m.p. 128°-129°; [α]_(D) =-50.1° (c=1, methanol).

Anal. calc'd. for C₁₂ H₁₄ O₃ S•C₁₀ H₁₅ NO: C, 65.48; H, 7.24; N, 3.47;S, 7.95 Found: C, 65.46; H, 7.34; N. 3.21; S, 8.00.

b)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[1,2-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A stirred suspension of the ephedrine salt from part (a) (333.1 mg.,0.822 mmol.) in ethyl acetate (5 ml) was washed three times with 5 mlportions of 1N hydrochloric acid solution. The organic extracts werecombined, washed with brine, dried (magnesium sulfate), filtered,concentrated and dried in vacuo for 30 minutes. The resulting oil wasdissolved in methylene chloride (2 ml) and stirred under nitrogen at 0°C. To this solution was added a solution of[4S-(4α,7α,10aβ)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylic acid, methyl ester [200.0 mg, 0.774mmol., prepared as described in Example 3(c)] in methylene chloride (6ml), then triethylamine (0.113 ml, 0.813 mmol ) and finallybenzotriazol-1-yloxtris (dimethylamino)phosphonium hexafluorophosphate(360.0 mg., 0.813 mmol.) The reaction was stirred at 0° C. and allowedto slowly warm to room temperature. After 19 hours, the reaction wasconcentrated in vacuo and the residue was dissolved in ethyl acetate.The solution was washed once with a 5% solution of potassium bisulfate(20 ml), once with a saturated solution of sodium bicarbonate (20 ml),and once with brine. The organic layer was dried (magnesium sulfate),filtered and concentrated to a yellow foam. Purification by flashchromatography (silica gel, 230-400 mesh under 10-20 psi of nitrogenpressure) eluting with 4:3 ethyl acetate/hexane gave 303 mg of productas a clear oil.

c)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-mercaptomethyl)-1-oxo-3-phenylpropyl]amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (b) (303.1 mg., 0.635 mmol) inmethanol (6.5 ml, deoxygenated via nitrogen bubbling) was cooled to 0°C. and treated with 1N sodium hydroxide (6.5 ml, deoxygenated vianitrogen bubbling). After stirring for one hour at 0° C. while purgingcontinuously with nitrogen, the reaction was warmed to room temperature.After a total of three hours, the reaction was acidified to pH 1 with 5%potassium bisulfate and extracted with ethyl acetate. The organic layerswere combined, washed with water, brine, dried (sodium sulfate),filtered, and concentrated in vacuo to give 219 mg of title product as awhite solid; m.p. 200° C. (decomp.). TLC (55 6:0.01:3.99 ethylacetate/acetic acid/hexane) R_(f) =0.15. HPLC: t_(R) =26.3 min, impurityat 27.0 min.; YMC S-3 ODS (C-18) 6.0×150 mm; 0% to 100% B: A, 30 min.linear gradient and 10 min. hold, 1.5 ml/min.; A=90% water:methanol+0.2%phosphoric acid, B=90% methanol:water+0.2% phosphoric acid; 220 nm.

Anal. calc'd for C₂₀ H₂₆ O₄ N₂ S₂ •0.11 C₄ H₈ O₂ •0.07 CH₂ Cl₂ : C,56.22; H, 6.21; N, 6.39; S, 14.63. Found: C, 56.46; H, 6.28; N, 6.31; S,14.59.

EXAMPLE 13

[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-4-methyl-1-oxopentyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

a) (R)-2-Bromo-4-methylpentanoic acid

Potassium bromide (9.5 g., 80 mmol) was added to a stirred solution ofD-leucine (3.0 g., 23 mmol) in 2.5N sulfuric acid (47 ml) at roomtemperature. The reaction mixture was cooled to -10° C. and solid sodiumnitrite (2.4 g., 34 mmol) was added portionwise, maintaining thetemperature between -10° and -5° C. After addition was complete, thereaction was stirred for 1 hour and then warmed to room temperature andstirred for another hour. The reaction mixture was then extracted twicewith ether, the ether extracts were washed once with water, dried(magnesium sulfate), filtered and evaporated to give 2.7 g of crudetitle product.

b) (S)-2-(Acetylthio)-4-methylpentanoic acid, dicyclohexylamine salt

To a stirred slurry of potassium thioacetate (1.7 g, 15.0 mmol) in 50 mlof dry acetonitrile at room temperature under argon was added a solutionof the product from part (a) (2.6 g., 13 mmol) in 17 ml of acetonitrile.The reaction was stirred 4 hours. The resulting slurry was filtered andevaporated. The residue was redissolved in ethyl ether, washed once with5% potassium bisulfate solution and once with brine, dried (magnesiumsulfate) and evaporated. The residue was dissolved in ether (64 ml) andtreated with dicyclohexylamine (2.7 ml, 14 mmol). A white solidimmediately began precipitating from the solution. The solution wasfiltered and the white solid collected to give 2.0 g of title product;m.p. 153°-158° C.; [α]_(D) =-54.5° (c=0.61, chloroform).

c)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-4-methyl-1-oxopentyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A stirred suspension of the product from part (b) (403.3 mg, 1.09 mmol)in ethyl acetate (5 ml) was washed three times with 5 ml portions of 5%potassium bisulfate solution. The organic extracts were combined, washedwith brine, dried (sodium sulfate), filtered, concentrated and dried invacuo for 30 minutes. The resulting oil (179.4 mg, 0.943 mmol) wasdissolved in methylene chloride (2 ml) and stirred under nitrogen at 0°C. To this solution was added a solution of[4S-(4α,7α,10aβ)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester [232.0 mg, 0.898 mmol., prepared as described inExample 3(c)] in methylene chloride (6 ml), then triethylamine (0.131ml., 0.943 mmol), and finallybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(417.1 mg, 0.943 mmol.). The reaction was stirred at 0° C. for one hourand 3.5 hours at room temperature. After a total of 4.5 hours, thereaction was concentrated in vacuo and the residue was dissolved inethyl acetate. The solution was washed once with a 5% solution ofpotassium bisulfate (20 ml), once with a saturated solution of sodiumbicarbonate (20 ml), and once with brine. The organic layer was dried(magnesium sulfate), filtered and concentrated to a yellow foam.Purification by flash chromatography (silica gel, 230-400 mesh lander10-20 psi of nitrogen pressure) eluting with 2:3 ethyl acetate/hexanegave 209.4 mg. of title product as a clear oil.

d)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-4-methyl-1-oxopentyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (c) (209.4 mg, 0.486 mmol) inmethanol (5 ml, deoxygenated via nitrogen bubbling) was cooled to 0° C.and treated with 1N sodium hydroxide (5 ml, deoxygenated via nitrogenbubbling). After stirring for one hour at 0° C. while purgingcontinuously with nitrogen, the reaction was warmed to room temperature.After a total of 2.5 hours, the reaction was acidified to pH 1 with 51%potassium bisulfate and extracted with ethyl acetate. The organic layerswere combined, washed with water, brine, dried (sodium sulfate),filtered, and concentrated in vacuo. Purification by flashchromatography (silica gel, 230-400 mesh under 10-20 psi of nitrogen)eluting with nitrogen sparged 6:0.01:3.99 ethyl acetate/aceticacid/hexane gave 142.0 mg of title product as a white solid. TLC(6:0.1:3.9 ethyl acetate/acetic acid/hexane) R_(f) =0.20. [α]_(D)=-103.0° (c=0.43, chloroform). HPLC: t_(R) 26.7 min.; YMC S-3 ODS (C-18)6.0×150 mm; 0% to 100% B: A, 30 min. linear gradient and 10 min. hold,1.5 ml/min; A=90% water:methanol+0.2% phosphoric acid, B=90%methanol:water+0.2% phosphoric acid; 220 nm.

Anal. calc'd for C₁₆ H₂₆ O₄ N₂ S₂ •0.19 C₄ H₈ O₂ •0.18 C₇ H₁₆ •0.9 H₂ O:C, 50.87; H, 7.63; N, 6.58; S, 15.07. Found: C, 50.57; H, 7.20; N, 6.83;S, 14.75.

EXAMPLE 14[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxybutyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

a) (R)-2-Bromobutanoic acid

Potassium bromide (7.85 g, 65.94 mmol) was added to a solution of(R)-2-aminobutanoic acid (2.0 g, 19.40 mmol) in 2.5N sulfuric acid (25ml) and cooled to 0° C. Sodium nitrite (2.06 g, 29.87 mmol) was addedslowly in several portions. The temperature was kept at less than 2° C.during this addition. The reaction was stirred at 0° C. for one hour andat room temperature for 16 hours, then extracted with 3-50 ml portionsof ethyl acetate. The combined ethyl acetate layers were washed with2-50 ml portions of water, 1-50 ml portion of brine, dried (magnesiumsulfate) and concentrated in vacuo to give 2.86 g of title product as acrude oil.

b) (S)-2-(Acetylthio)butanoic acid, dicyclohexylamine salt

To a slurry of 2.14 g (18.77 mmol) of potassium thioacetate in 15 ml ofacetonitrile, stirred at room temperature, was added dropwise over 15minutes a solution of (R)-2-bromobutanoic acid (2.83 g, 17.07 mmol) in15 ml of acetonitrile. The reaction was stirred at room temperature for16 hours then filtered. The filtrate was concentrated in vacuo and theresulting oil was dissolved in 30 ml of ethyl ether. The ether layer waswashed with 2-20 ml portions of 5% potassium bisulfate, 2-20 ml portionsof water, 2-20 ml portions of brine, dried (magnesium sulfate) andfiltered. To the filtrate was added 3.4 ml (17.07 mmol) ofdicyclohexylamine. After stirring at room temperature for 2 hours, theslurry was filtered to give 1.24 g of the dicyclohexylamine saltproduct. A second crop of 724 mg of dicyclohexylamine salt product wasobtained from the filtrate.

c)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-[(acetylthio)-1-oxobutyl]amino]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid, methyl ester

The dicyclohexylamine salt product from part (b) (227 mg, 0.66 mmol) wasdissolved in 15 ml of ethyl acetate and washed with three 10 ml portionsof potassium bisulfate, one 10 ml portion of brine, dried (magnesiumsulfate), and concentrated in vacuo to give 108 mg of(S)-2-(acetylthio)butanoic acid as a clear oil.

To a solution of this free acid (108 mg, 0.66 mmol) in 5 ml of drymethylene chloride, cooled to 0° C., was added triethylamine (90 μl,0.66 mmol), followed by[4S-(4α,7α,9aβ)]-4-amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [250.0 mg, 0.66 mmol,prepared from the material described in Example 5(d)] and a secondportion of triethylamine (90 μl, 0.66 mmol). The reaction was stirred at0° C. for 20 minutes, thenbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(292 mg, 0.66 mmol) was added in one portion. The reaction was stirredat 0° C. for one hour, refrigerated for 56 hours, then stirred at roomtemperature for 3 hours. The reaction mixture was then concentrated invacuo, redissolved in 30 ml ethyl acetate and washed with 20 ml of 5%potassium bisulfate, 20 ml saturated sodium bicarbonate, 20 ml of brine,dried (magnesium sulfate), and concentrated in vacuo to give a crudeoil. The crude oil was flash chromatographed (Merck silica gel, 25×100mm, 2:3 ethyl acetate/hexane) to give 208 mg of title product as a whitefoam.

d)4S-4α(R*),7α,9aβ]]-OCTAHYDRO-4-[(2-mercapto-1-oxybutyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (c) (225 mg, 0.559 mmol) in methanol(5 ml) was purged with argon for 30 minutes and cooled to 0° C. To thissolution was added dropwise 5 ml of 1M sodium hydroxide, also purgedwith argon for 30 minutes and cooled to 0° C. The reaction was stirredat 0° C. for 3 hours with continuous argon purging, then acidified to pH2 with 5% potassium bisulfate solution. The mixture was extracted with3-40 ml portions of ethyl acetate, and the combined ethyl acetate layerswere dried (magnesium sulfate), and concentrated in vacuo to give acrude foam. The crude product was flash chromatographed (Merck silicagel, 25×180 mm, 3% acetic acid/ethyl acetate) to give a white foam,which was dissolved in methylene chloride and triturated with hexane togive 176 mg of title product as a compact white foam; [α]_(D) =-125.4°(c=1.0, chloroform). TLC (methanol/methylene chloride 1:9) R_(f) =0.16.HPLC: t_(R) =15.5 min. (97% total area, UV 220 nM); YMC S-3 ODS (C-18,120A) 6×150 mm; 0% B:A-100% B:A, linear 25 minute gradient (A=90%water/methanol+0.2% phosphoric acid) B=90% methanol/water+0.2%phosphoric acid); flow rate =1.5 ml/min.

Anal. calc'd for C₁₃ H₂₀ N₂ S₂ O₄ •0.8 H₂ O•0.2 C₆ H₁₄ •0.1 CH₂ Cl₂ : C,46.10; H, 6.66; N, 7.52; S, 17.21. Found: C, 46.00; H, 6.17; N, 7.52; S,16.82.

EXAMPLE 15[4S-[4α(R*),7α,9aβ]]-(Octahydro-4-[(2-mercapto-1-oxopentyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

a) (R)-2-Bromopentanoic acid

Following the procedure of Example 14(a) but employing D-norvaline inplace of (R)-2-aminobutanoic acid, (R)-2-bromopentanoic acid wasobtained as a clear liquid.

b) (S)-2-(Acetylthio)pentanoic acid, dicyclohexylamine salt

Reacting (R)-2-bromopentanoic acid with potassium thioacetate inacetonitrile followed by treatment with dicyclohexylamine according tothe procedure of Example 14(b), (S)-2-(acetylthio)pentanoic acid,dicyclohexylamine salt was obtained as a white solid.

c)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxopentyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

Reacting the free acid of the dicyclohexylamine salt from part (b) with[4S-[(4α,7α,9aβ)]4-amino-octahydro-5-oxopyrrolo-[2,1b][1,3]thiazepine-7-carboxylicacid, methyl ester according to the procedure of xample 14(c), the titleproduct was obtained as a white foam.

d)[4S-[4α(R*),7α,9aα]]-Octahydro-4-[(2-mercapto-1-oxopentyl)amino]-5-oxopyrrolo-[2,1-b][1,3]thiazpine-7-carboxylicacid

A solution of the product from part (c) in methanol was treated with 1Msodium hydroxide according to the procedure of Example 14(d) and gavethe title product as a compact white foam; [α]_(D) =-122.8° (c=1.0,CDCl₃). TLC (methanol/methylene chloride 1:9) R_(f) =0.15. HPLC: t_(r)=20.5 min.; (97% total area, UV 220 nM); YMC S-3 ODS (C-18, 120A) 6×150mm; 0% B:A- 100% B: A, linear 25 minute gradient (A=90%water:methanol+0.2% phosphoric acid); B=90% methanol:water+0.2%phosphoric acid; flow rate=1.5 ml/min.

Anal. calc'd for C₁₄ H₂₂ N₂ S₂ O₄ •0.65 H₂ O•0.20 C₆ H₁₄ : C, 48.63; H,7.01; N, 7.46; S, 17.08. Found: C, 48.86; H, 6.70; N, 7.24; S, 16.74.

EXAMPLE 16 [4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-4,4-dimethyl-1-oxopentyl)]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid

a) (R) 2-Bromo-4,4-dimethylpentanoic acid

A solution of (R)-2-amino-4,4-dimethylpentanoic acid (950 mg, 6.55 mmol)in 2.5N aqueous sulfuric acid (13 ml, 33 mmol) was cooled to -5° C. andtreated with potassium bromide (2.72 g, 22.9 mmol) in one portion. Thecolorless solution was treated with sodium nitrite (680 mg, 9.86 mmol)portionwise, keeping the temperature between 0° and 3° C. over a periodof 25 minutes. The reaction mixture was stirred at 0° C. for one hourand at room temperature for 1.5 hours. The reaction mixture was pouredinto water (10 ml). The product was extracted with ether (60 ml), washedwith water (20 ml) and brine (20 ml), dried (magnesium sulfate), andconcentrated in vacuo to give the title product as a colorless liquid.

b) (S)-2-(Acetylthio)-4,4-dimethylpentanoic acid

A slurry of potassium thioacetate (750 mg, 6.58 mmol) and acetonitrile(10 ml, molecular sieves dried) was cooled to 0° C. and treated with asolution of (R)-2-bromo-4,4-dimethylpentanoic acid from part (a) inacetonitrile (2 ml) over 5 minutes. The reaction mixture was allowed towarm to room temperature and stirred for 2.5 hours. The slurry wasfiltered. The filtrate was concentrated in vacuo. The residue wasdiluted in ether (50 ml), washed with two portions of 5% aqueous sodiumthiosulfate (50 ml) and brine (25 ml), dried (magnesium sulfate), andconcentrated in vacuo. The pale yellow oil was purified by flashchromatography (Merck silica gel, 12×3 cm., 10% methanol/methylenechloride) affording 575 mg of title product as a pale yellow oil.

c)[4S-[4α(R*),7α,9aβ]]-OCTAHYDRO-4-[[2-(acetylthio)-4,4-dimethyl-1-oxopentyl]amino]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid, methyl ester

A clear solution of the product from part (b) (148 mg, 0.72 mmol) inmethylene chloride (5 ml, distilled from calcium hydride) was cooled to0° C. and treated with a solution of[4S-(4α,7α,9aβ)]-4-amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [250.0 mg, 0.60 mmol,prepared from the material described in Example 5(d)] in methylenechloride (3 ml, distilled from calcium hydroxide), triethylamine (122mg, 1.2 mmol), followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(319 mg, 0.72 mmol). The reaction mixture was stirred at 0° C. for 24hours and at room temperature for 5 hours. The crude reaction mixturewas concentrated in vacuo. The residue was diluted in ethyl acetate (50ml), washed with 5% aqueous potassium bisulfate (50 ml), 50% saturatedaqueous sodium bicarbonate solution (50 ml), and brine (50 ml), dried(sodium sulfate) and concentrated in vacuo. The crude product waspurified by flash chromatography (20 g, Merck silica gel, ethyl acetate)to afford 244 mg of the title product as a white foam.

d)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-4,4-dimethyl-1-oxopentyl)amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid

A clear solution of the product from part (c) (240 mg, 0.56 mmol) inmethanol (2 ml, nitrogen sparged) was cooled to 0° C. and treateddropwise with 1N sodium hydroxide (2.27 ml, 2.24 mmol, argon sparged)with continuous sparging at 0° C. The mixture was allowed to stir at 0°C. for 3 hours and at room temperature for 3 hours. The mixture wasacidified to pH of 1 with a solution of 5% aqueous potassium bisulfate(argon sparged). The product was extracted with methylene chloride (50ml, nitrogen sparged), washed with brine, dried (sodium sulfate) andconcentrated in vacuo. The crude product was recrystallized frommethylene chloride/hexane to afford 75 mg of title product as a whitesolid; m.p. 124°-126° C.; [α]_(D) =-175° (c=0.25, methanol). TLC (aceticacid/methanol/methylene chloride 1:5:94)R_(f) =0.65. HPLC: t_(R) (YMC,S-3 ODS (C-18) 6.0×150 mm; 1.5 ml/min. linear gradient 0-100%B over 30min., Buffer A=methanol/water/phosphoric acid (10:90:0.2), BufferB=methanol/water/phosphoric acid (90:10:0.2))=24.4 min., 95% of totalpeak area at 254 nm.

Analysis calc'd. for C₁₆ H₂₆ N₂ O₄ S₂ •0.13 C₆ H₁₄ : C, 52.24; H, 7.28;N, 7.26; S 16.62 Found: C, 51.97; H, 7.26; N, 7.09; S, 16.23.

EXAMPLE 17[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b]1,3]thiazepine-7-carboxylicacid

a) [(R)-2-Bromopropanoic acid

Following the procedure of Example 14(a) but employing D-alanine inplace of (R)-aminobutanoic acid, (R)-2-bromopropanoic acid was obtainedas a light yellow oil.

b) (S)-2-(Acetylthio)propanoic acid

To a light green solution of potassium thioacetate (3.94 g, 34.5 mmol)in acetonitrile (150 ml) was added a solution of (R)-2-bromopropanoicacid (4.8 g, 31 mmol) in acetonitrile (12 ml) at room temperature underan argon atomosphere. The resulting white slurry was stirred at roomtemperature for 2 hours then filtered. The filtrate was concentrated invacuo. The residue was diluted in ethyl acetate (100 ml), washed with a10% aqueous solution of potassium bisulfate (50 ml) and brine, dried(sodium sulfate), and concentrated in vacuo. The crude product (4.61 g)was purified by flash chromatography (60 g-Merck silica gel, 1:45:54acetic acid/ethyl acetate/hexane) to afford 3.7 g of the title productas a light yellow oil; [α]_(D) =-114° (c=0.50, methanol).

c)[4S-4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxopropyl]amino]-5-oxopyrrolo[2,1-b][1,3-]thiazepine-7-carboxylicacid, methyl ester

A clear solution of (S)-2-(acetylthio)propanoic acid (86 mg, 0.58 mmol)in methylene chloride (5 ml, distilled from calcium hydride) was cooledto 0° C. and treated with a solution[4S-(4α,7α,9aβ)]-4-amino-octahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [200.0 mg, 0.48 mmol,prepared from the material described in Example 5(d)]in methylenechloride (5 ml, distilled from calcium hydride), triethylamine (98 mg,0.97 mmol), followed bybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(255 mg, 0.58 mmol). The reaction mixture was stirred at 0° C. for 22hours and at room temperature for 2 hours. The crude reaction mixturewas concentrated in vacuo. The residue was diluted in ethyl acetate (100ml), washed with 5% aqueous potassium bisulfate (30 ml), 50% saturatedaqueous sodium bicarbonate solution, and brine, dried (sodium sulfate),and concentrated in vacuo. The crude product was purified by flashchromatography (40 g., Merck silica gel, ethyl acetate) to afford 180 mgof title product as a white solid; m.p. 143°-145° C.

d)[4S-4α(R*),7α,9aβ]]-Octahydro-4-[(2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b[1,3]thiazepine-7-carboxylicacid

A clear solution of the product from part (c) (180 mg, 0.48 mmol) inmethanol (2 ml) under an argon atmosphere was cooled to -10° C. andtreated dropwise with argon sparged 1N sodium hydroxide (1.95 ml, 1.95mmol), keeping the temperature below 0° C. The mixture was allowed tostir with argon sparging at 0° C. for 3 hours. The mixture was acidifiedto pH of 1 with a solution of 5% aqueous potassium bisulfate under anargon atmosphere. The product was extracted with nitrogen sparged ethylacetate (100 ml), washed with brine, dried (sodium sulfate), andconcentrated in vacuo. The crude product was purified by flashchromatography (40 g, Merck silica gel, 1:5:94 aceticacid/methanol/methylene chloride) to afford 154 mg of title product as awhite solid; m.p. 150°-152° C.; [α]_(D) =-156° (C=0.50, methanol). TLC(1:5:94 acetic acid/methanol/methylene chloride) R_(f) =0.28. HPLC:t_(R) (YMC, S-3 ODS (C-18) 6.0×150 mm; 1.5 ml/min. linear gradient0-100% B over 30 minutes. Buffer A=methanol/water/phosphoric acid(10:90:0.2), Buffer B=methanol/water/phosphoric acid (90:10:0.2))=14.69min., more than 95% of total peak area at 254 mM.

Anal. calc'd. for C₁₂ H₁₈ N₂ O₄ S₂ •0.75CH₃ CO₂ H: C, 44.61; H, 5.82; N,7.71; S, 17.64 Found: C, 44.76; H, 5.71; N, 7.81; S, 17.76.

EXAMPLE 18[4S-[4α(R*),7α,9aβ]]-Octahydro-4-cyclopropyl-2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b][1,3]-thiazepine-7-carboxylicacid

a) (R)-2-[[Phenylmethoxy)carbonyl]amino]-4-pentenoic acid

A mixture of D-allylglycine (2.8 g, 24.3 mmol), 1M aqueous sodiumhydroxide solution (25 ml), and tetrahydrofuran (10 ml, distilled fromketyl) was stirred at room temperature until homogeneous then cooled inan ice-bath. To the resulting rapidly stirred solution was added about 5mL of 1.0M aqueous sodium hydroxide solution then dropwise about 1 g ofbenzyl chloroformate. This was repeated 4 additional times until a totalof 28 mL of 1.0M aqueous sodium hydroxide solution and 4.80 g (95%, 27mmol) of benzyl chloroformate were added. The reaction mixture wasstirred for 15 minutes at 0° C. then 30 minutes at room temperature andthen extracted with 50 mL of ether. The aqueous layer was acidified(pH=1.5) the by addition of 6N hydrochloric acid solution (about 10 mL)then extracted with three-50 mL portions of ether. The three etherextracts were combined, dried (magnesium sulfate) and concentrated invacuo to afford 6.01 g of title product as a colorless oil.

b) (R)-2-[[(Phenylmethoxy)carbonyl]amino]-4-pentenoic acid, phenylmethylester

Cesium carbonate (4.28 g, 13.1 mmol) was added to a solution of theproduct from part (a) (5.96 g, 23.9 mmol) in anhydrous dimethylformamide(25 ml) at room temperature. The reaction mixture was stirred for 20minutes then benzyl bromide (4.5 g, 26.3 mmol) was added rapidly (mildlyexothermic). The mixture was stirred for 30 minutes then partitionedbetween 100 ml of water and 100 ml of ether. The organic layer wasseparated, washed with three-100 mL portions of water, 50 mL of brine,dried (magnesium sulfate) and concentrated in vacuo to give an oil. Thecrude material was purified by flash chromatography (Merck silica gel,24×5.0 cm, 1:10 ethyl acetate/hexane then 1:4 ethyl acetate/hexane) toafford 6.13 g of title product as a colorless oil.

c)(R)-α-[[(Phenylmethoxy)carbonyl]amino]cyclopropanepropanic acid,phenylmethyl ester

Palladium(II)acetate (65 mg, 0.29 mmol) was added to a solution of theproduct from part (b) (5.78 g, 17.1 mmol) in anhydrous ether (60 ml) andstirred for 10 minutes. The resulting mixture was cooled to 0° C. wasexcess ethereal diazomethane (prepared from 12 gN-methyl-N'-nitro-N-nitrosoguanidine/120 ml ether) was added in portionsover about 15 minutes. The reaction mixture was stirred for 15 minutesthen quenched by addition of 1 mL of glacial acetic acid. The solutionwas transferred to a separatory funnel, washed with 100 mL of saturatedaqueous sodium bicarbonate solution, 50 mL of brine, dried (magnesiumsulfate) and concentrated in vacuo to give a yellow oil. The crudematerial was purified by flash chromatography (Merck silica gel, 20×5.0cm, 1:4 ethyl acetate/hexane) to afford 5.74 g of title product as acolorless oil.

d) (R)-(α-Amino)cyclopropanepropanoic acid

Palladium on carbon catalyst (10%, 1.14 g) was added to a solution ofthe product from part (c) (5.71 g, 16.2 mmol) in methanol (75 ml) andstirred under an atmosphere of hydrogen (balloon) at room temperaturefor 48 hours. The reaction mixture was filtered to remove the catalystand the catalyst was rinsed with warm water. The filtrate was thenpassed through a 0.4 μM polycarbonate membrane filter. The filtrate wasconcentrated in vacuo to give 2.03 g of title product as a white solid.

e) (S)-α-(Acetylthio)cyclopropanepropanoic acid

A mixture of the product from part (d) (2.00 g, 15.5 mmol) in 2.5Naqueous sulfuric acid (30 ml) was stirred at room temperature untilhomogeneous and then cooled to -5° C. Potassium bromide (6.50 g, 54.6mmol) was added to this solution in one portion. The mixture was stirreduntil homgeneous. Sodium nitrite (1.60 g, 23.2 mmol) was then added insmall portions over about 25 minutes, maintaining the reactiontemperature below 0° C. The reaction mixture was stirred for anadditional 1 hour at 0° C. then at room temperature for 1.5 hours. Theresulting mixture was diluted with 30 ml of water and extracted withthree-30 mL portions of ether. The combined ether extracts were washedwith 25 mL of brine, dried (magnesium sulfate) and concentrated in vacuoto give 2.82 g of crude (R)-(α-bromo)cyclopropanepropanoic acid as apale yellow oil.

A solution of this crude (R)-(α-bromo)cyclopropanepropanoic acid wasadded in acetonitrile (10 ml) over 5 minutes to a stirred slurry ofpotassium thioacetate (1.83 g, 16.1 mmol) in acetonitrile (20 ml) whilecooling in an ice-bath. The reaction mixture was stirred at 0° C. for 1hour then at room temperature for 16 hours. The reaction mixture wasfiltered and the filtrate concentrated in vacuo to give an oil. The oilwas partitioned between 50 mL of ether and 50 mL of 5% aqueous sodiumthiosulfate solution. The organic layer was separated, washed with 25 mLof brine, dried (magnesium sulfate) and concentrated in vacuo to give ayellow oil. The crude material was purified by flash chromatography(Merck silica gel, 12×5.0 cm, 1:19 methanol/methylene chloride) toafford 1.52 g of title product as a yellow oil.

f)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[[2-(acetylthio)-3-cyclopropopyl-1-oxopropyl]amino]-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(292 mg, 0.66 mmol) was added in one portion to a mixture of[4S-(4α,7α,9aβ)]-octahydro-4-amino-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [250 mg, 0.60 mmol,prepared from the material described in Example 5(d)] in methylenechloride (3 ml, distilled from calcium hydride), triethylamine (121 mg,1.20 mmol), and the product from part (e) (122 mg, 0.65 mmol) inmethylene chloride (3 ml, distilled from calcium hydride). The reactionmixture was stirred for 0° C. for 1 hour then at room temperature for1.5 hours. The resulting mixture was partitioned between 20 ml of ethylacetate and 20 ml of 1M aqueous potassium bisulfate solution. Theorganic layer was separated, washed with 20 ml of 5% aqueous sodiumbicarbonate solution, 20 ml of brine, dried (magnesium sulfate) andconcentrated in vacuo to give an oil. The crude material was purified byflash chromatography (Merck silica gel, 12×3.0 cm, 1:1 ethylacetate/hexane) to afford 191 mg of title product as a solid white foam.

g)[4S-[4α(R*),7α,9aβ]]-Octahydro-4-[(3-cyclopropropyl-2-mercapto-1-oxopropyl)amino]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid

A solution of the product from part (f) (185 mg, 0.45 mmol) in methanol(3 ml) was sparged with argon for 10 minutes at 0° C. and then 1Maqueous sodium hydroxide solution (3 ml, freshly sparged with argon for10 minutes) was added. The reaction mixture was stirred for 2.5 hours atroom temperature with continuous argon sparge then acidified by additionof 20 ml of 1M potassium bisulfate solution and extracted with 20 ml ofethyl acetate. The organic extract was washed with 20 ml of brine, dried(sodium sulfate) and concentrated in vacuo to give a gum. The gum wasrinsed with anhydrous ether then concentrated under oil pump vacuum toafford 121 mg of the title product as a white foam; [α]_(D) =-103°(c=0.23, methanol). TLC (1:10:90 acetic acid/methanol/methylenechloride) R_(f) =0.46. HPLC: t_(R) (YMC S-3 ODS 6.0×150 mm; 1.5 mL/min,linear gradient 0-100% B over 30 minutes, BufferA=methanol/water/phosphoric acid (10:90:0.2), BufferB=methanol/water/phosphoric acid (90:10:0.2))=20.7 minutes greater than97% of total peak area at 254 nm.

Anal. calc'd for C₁₅ H₂₂ N₂ O₄ S₂ •0.20 H₂ O: C, 49.77; H, 6.23; N,7.74; S, 17.71 Found C, 50.01; H, 6.27; N, 7.50; S, 17.40.

EXAMPLE 19

a)[4S-(4α,7α,9aβ-Octahydro-4-[[(1-meraptocyclopentyl)carbonyl]amino]-5-oxopyrrolo[2,1-b]-[1,3]thiazepine-7-carboxylicacid

a) 1-Mercaptocyclopentanecarboxylic acid

A solution of lithium diisopropylamide was prepared under nitrogen fromdiisopropylamine (5.4 ml, 38.5 mmol) and n-butyllithium (2.5M inhexanes, 15.4 ml, 38.5 mmol) in tetrahydrofuran (17.6 ml), maintainingthe temperature between -3° C. to 0°. After stirring for 15 minutes,cyclopentanecarboxylic acid (2.0 g, 17.5 mmol) was added intetrahydrofuran (2 ml) at 0° C. to 3° C. over 25 minutes. After 15minutes at 0° C., the bath was removed and the reaction was stirred 15minutes more, causing the temperature to rise to 15° C. The milky whitesolution was cooled to -78° C. and sulfur (S₈, 618.0 mg, 19.3 mmol) wasadded as a solid, maintaining the temperature at -78° C. The reactionwas allowed to warm to room temperature in situ. After 70 hours, thereaction was cooled to 0° C., quenched with water (pH 8-9) and quicklyacidified to pH 1 with 6N hydrochloric acid. The aqueous solution wasextracted with ethyl acetate (3×30 ml), washed with brine, dried(magnesium sulfate), filtered and concentrated to give 2.62 g of titleproduct as a yellow oil.

b) 1-(Acetylthio)cyclopentanecarboxylic acid

To a solution of the product from part (a) (1.44 g, 9.89 mmol) in anitrogen sparged solution of 1N sodium hydroxide (20 ml, 19.7 mmol) at0° C. was added acetic anhydride (0.93 ml, 9.89 mmol). Tetrahydrofuran(13 ml) was added in order to solubilize the oil which formed. Afterstirring one hour at 0° C. (pH 7), the reaction was warmed to roomtemperature and additional acetic anhydride (0.47 ml, 4.9 mmol) wasadded, as well as solid potassium carbonate (2.04 g, 14.8 mmol) to pH 10and tetrahydrofuran (4 ml). After stirring overnight at roomtemperature, the reaction mixture was acidified to pH 1 with 1Nhydrochloric acid and extracted with ethyl acetate. The ethyl acetateextracts were combined, washed with brine, dried (magnesium sulfate)filtered and concentrated in vacuo to give a yellow solid (1.61 g). Thesolid was recrystallized twice from ethyl acetate/hexanes to give 614 mgof title product as a light brown solid; m.p. 119.5°-121.5° C.

c)[4S-(4α,7α,9aβ)]-Octahydro-4-[[1-[(acetylthio)cyclopentyl]carbonyl]amino-5-oxopyrrolo[2,1-b][1,3]-thiazepine-7-carboxylicacid, methyl ester

To a solution of the product from part (b) (94.5, 0.502 mmol) inmethylene chloride (3.6 ml) at 0° C. under nitrogen, was addedtriethylamine (70 μl, 0.502 mmol) followed by[4S-(4α,7α,9aβ)]-4-aminooctahydro-5-oxopyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt [prepared from thematerial described in Example 5(d), 198.9 mg, 0.478 mmol] in oneportion, followed by triethylamine (66.6 μl, 0.478 mmol). The reactionwas stirred for 5 minutes at 0° C.Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(222.0 mg, 0.502 mmol) was then added as a solid. The reaction wasstirred at 0° C. for one hour and then at room temperature for 2.25hours. The reaction was concentrated in vacuo and the residuepartitioned between ethyl acetate and 5% potassium bisulfate (20 ml).The organic layer was washed with half saturated sodium bicarbonate andbrine, dried (magnesium sulfate), filtered and concentrated to a clearoil. Purification by flash chromatography eluting with 11:9 ethylacetate/hexane gave 169.3 mg. of title product as a clear oil.

d)[4S-(4α,7α,9aβ)]-Octahydro-4-[[(1-mercaptocyclopentyl)carbonyl]amino]-5-oxopyrrolo[2,1-b][1,3]-thiazepine-7-carboxylicacid

A solution of the product from part (c) (167.3 mg., 0.404 mmol) inmethanol (4 ml, deoxygenated via nitrogen bubbling) was cooled to 0° C.and treated with 1N sodium hydroxide (4 ml, deoxygenated via nitrogenbubbling). After stirring for 1.5 hours at 0° C. while purgingcontinuously with nitrogen, the reaction was warmed to room temperature.After a total of three hours, the reaction was acidified to pH 1 with 5%potassium bisulfate and extracted with ethyl acetate. The organic layerswere combined, washed with water (20 ml), brine, dried (sodium sulfate),filtered, concentrated in vacuo and re-evaporated from hexanes to give awhite solid. The compound was dissolved in dioxane (anhydrous) andlyophillized to give 110 mg, of title product as a white solid; [α]_(D)=-106.5° (c=0.68, chloroform). TLC (7:2.9:0.1, ethylacetate/hexane/acetic acid)R_(f) =0.12. HPLC: t_(R) =21.5 min; YMC S-3ODS (C-18) 6.0×150 mm; 0% to 100% B: A, 30 minutes linear gradient and10 minutes hold, 1.5 ml/min.; A=90% water/10% methanol+0.2% phosphoricacid, B=90% methanol/10% water+0.2% phosphoric acid; 220 nm.

Anal. calc'd for C₁₅ H₂₂ N₂ O₄ S₂ •0.15 C₄ H₈ O₂ •0.7 H₂ O•0.08C₆ H₁₄ :C, 49.37; H, 6.63; N, 7.16; S, 16.39 Found: C, 49.03; H, 6.37; N, 7.21;S, 16.65.

EXAMPLE 204S-(4α,7α,10β)]-4-[(2-Carboxy-1-oxo-3-phenylpropyl)amino]octahydro-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylicacid

3-(Phenylmethyl)propanedioic acid, monoethyl ester

a) 3-(Phenylmethyl)propanedioic acid, diethyl ester (2.5 g, 10 mmol) in10 ml of tetrahydrofuran was stirred overnight with 10 ml of 1N lithiumhydroxide. The reaction mixture was acidified with 11 ml of 1Nhydrochloric acid and extracted with two 50 ml portions of ethylacetate. The ethyl acetate extracts were washed with brine, dried(sodium sulfate), and concentrated in vacuo. The concentrate waschromatographed through silica gel (80 g) using a 5% methanol:chloroformsolvent system. The appropriate fractions were combined and concentratedto yield 1.23 g of title product.

b)4S-(4α,7α,10aβ)]-4-(Ethoxycarbonyl)-1-oxo-3-phenylpropyl)amino]octahydro-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylicacid, methyl ester

The product from part (a) (0.222 g, 1 mmol) and[4S-(4α,7α,10aβ)-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester [0.258 g., 1 mmol, prepared as described in Example3(c)] were dissolved in methylene chloride (5 ml) and cooled to 0° C.Triethylamine (0.14 ml, 1 mmol) was added and the reaction mixture wasstirred for one hour. Benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (0.442 g, 1 mmol) was added and the solution wasstirred at 0° C. for 1 hour and at room temperature for 2.5 hours. Thereaction mixture was diluted with 50 ml of methylene chloride and washedwith water, 10% sodium bisulfate, saturated aqueous sodium bicarbonate,dried (sodium sulfate), filtered and concentrated in vacuo. The residuewas chromatogaphed through silica gel using 30% ethyl acetate inhexanes. The appropriate fractions were combined and concentrated invacuo to yield 0.22 g of title product.

c)4S-(4α,7α,10aβ)]-4-[(2-Carboxy-1-oxo-3-phenylpropyl)amino]octahydro-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylicacid

The product from part (b) (0.22 g, 0.476 mmol) was stirred with 1Nlithium hydroxide (5 ml) in tetrahydrofuran (5 ml) at room temperaturefor 3 hours. The reaction mixture was acidified to pH 2 with 1Nhydrochloric acid and concentrated in vacuo. The residue was dissolvedin ethyl acetate and washed with water, brine, dried (sodium sulfate),and concentrated in vacuo to 3 ml at which point product crystallized.After standing at 0° C. overnight, the solid was filtered and dried toyield 0.16 g of title product as a white solid; m.p. 159°-162° C.;[α]_(D) =-84.92° (c =0.7, methanol). TLC (chloroform:methanol, 9:1)R_(f) =0.23, 0.28. HPLC: t_(R) =16.15, 16.35 min.; (UV 254 nm); YMC S-3ODS (C-18) 6.0×150 mm, 3μ end capped column, linear gradient of 50-90%aqueous methanol containing 0.2% phosphoric acid, 20 min., 1.5 ml/min.(44.9%, 55.1% isomer mixture).

Anal. calc'd for C₂₀ H₂₄ N₂ SO₆ •0.1 H₂ O: C, 56.89; H, 5.78; N, 6.66;S, 7.59 Found: C, 56.98; H, 5.68; N, 6.58; S, 7.15.

EXAMPLE 21[4S-4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo[1,4]oxazino[3,4-b][1,3]thiazepine-7-carboxylic acid

a)O-(2,2-Dimethoxyethyl)-N-[N-[(phenylmethoxy)carbonyl]-L-homoseryl]-L-serine,methyl ester

A solution ofN-[O-[(1,1-dimethylethyl)dimethylsilyl]-N-[(pheenylmethoxy)carbonyl]-homoseryl]-O-)2,2-dimethoxyethyl)-L-serine,methyl ester [5.56 g, 10 mmol, prepared as described in Example 10(h)]in methanol (65 ml) was cooled to 0° C. (ice salt bath), treated withp-toluenesulfonic acid monohydrate (386 mg, 2.0 mmol) and stirred at 0°C. for 1.5 hours. The reaction was quenched with sodium bicarbonatesolution (198 mg. in 20 ml water), stirred for 5 minutes then evaporatedto remove the methanol. The aqueous phase was extracted with ethylacetate (2×200 ml) and the combined organic extract was washed withwater (110 ml), 5% sodium bicarbonate (80 ml) and brine (80 ml), dried(anhydrous sodium sulfate), filtered, evaporated to dryness and dried invacuo. The crude product was chromatographed on silica gel column(Merck), eluting the column with ethyl acetate:hexane (2:1) and ethylacetate:methanol (98:2) to give 3.975 g of title product as a syrup. TLC(ethyl acetate:hexane, 4:1) R_(f) =0.17.

b)O-(2,2-Dimethoxyethyl)-N-[O-(methylsulfonyl)-N-[(phenylmethoxy)carbonyl]-L-homoseryl]-L-serine,methyl ester

A solution of the product from part (a) (3.975 g, 8.98 mmol) in drymethylene chloride (52 ml) was cooled to -15° C., treated withtriethylamine (1.82 ml, 13.1 mmoles) and methanesulfonyl chloride (0.82ml, 10.6 mmoles) and stirred at -15° C. for 30 minutes. The reactionmixture was quenched with 25% ammonium chloride (19 ml), warmed to roomtemperature and diluted with ethyl acetate (750 ml). The organic phasewas washed with 5% potassium bisulfate (100 ml), 50% saturated brine(100 ml) and saturated brine (100 ml), dried (anhydrous sodium sulfate),filtered, evaporated to dryness and dried in vacuo to give 4.9 g oftitle product as a waxy solid. TLC (ethyl acetate:hexane, 4:1) R_(f)=0.32.

c)N-[S-Acetyl-N-[(phenylmethoxy)carbonyl]-L-homocysteinyl]-O-(2,2-dimethoxyethyl)-L-serine,methyl ester

Cesium carbonate (5.56 g, 17.04 mmoles) was added to a solution ofthiolacetic acid (2.6 ml) in dry methanol (40 ml), stirred for 10minutes then evaporated to dryness. The resulting solid was trituratedwith acetone (7×8 ml) and the off-white solids obtained were dried invacuo to give 4.39 g cesium thiolacetic acid.

A suspension of cesium thiolacetic acid (2.43 g, 1.3 eq.) in drydimethylformamide (8.0 ml) was treated with a solution of the productfrom part (b) (4.9 g, 8.98 mmol) in dry dimethylformamide (24 ml) andstirred for 16 hours at room temperature under argon. The mixture wasdiluted with ethyl acetate (1.0 L), washed successively with 5% sodiumbicarbonate (2×150 ml), water (2×150 ml) and brine (150 ml), dried(anhydrous sodium sulfate), filtered, evaporated to dryness and dried invacuo. The crude product was chromatographed on a silica gel column(Merck), eluting the column with ethyl acetate:hexane mixtures (1:1;2:1) to give 3.93 g of the title product as a waxy solid. TLC (ethylacetate:hexane, 4:1) R_(f) =0.63.

d)O-(2,2-Dimethoxyethyl)-N-[N-[(phenylmethoxy)carbonyl]-L-homocysteinyl]-L-serine,methyl ester

A solution of the product from part (c) (200 mg, 0.49 mmol) in methanol(8.0 ml) was purged with argon for 30 minutes, cooled to 0° C. (ice-saltbath) and treated with 25% sodium methoxide in methanol (0.11 ml, 0.5mmol), maintaining the bubbling of argon throughout the addition andlength of the reaction. After 5 minutes at 0° C., the mixture wasquenched with 25% ammonium chloride (2.3 ml) and partitioned betweenethyl acetate (2×12 ml) and water (2.3 ml). The combined organicextracts were washed with 25% ammonium chloride (4.6 ml) and brine (4.6ml), dried (anhydrous sodium sulfate), filtered, evaporated to drynessand dried in vacuo to give 183.2 mg of title product as a white solid.TLC (ethyl acetate:hexane, 4:l)R_(f) =0.62.

e)[4S-(4α,7α,10aβ)]-Octahydro-5-oxo-4-[[(phenylmethoxy)carbonyl]amino][1,4]oxazino[3,4-b][1,3]-thiazepine-7-carboxylicacid

A solution of the product from part (d) (50 mg, 0.11 mmol) in drymethylene chloride (2.0 ml) was treated with Amberlyst® 15 (H⁺ form; 13mg), stirred for 3 days at room temperature under argon, treated withmore Amberlyst® 15 (13 mg) and stirred for another 3 days. The solutionwas decanted and chromatographed on silica gel column (Merck), elutingthe column with ethyl acetate:hexane mixtures (1:3; 1:1) to give 21.1 mgof title product as a syrup. TLC (ethyl acetate:hexane, 4:1) R_(f)=0.70.

f)[4S-[4α,7α,10aβ)]-Octahydro-4-amino-5-oxo[1,4]-oxazino[3,4-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of the product from part (e) (421 mg, 1.01 mmoles) in drymethylene chloride (25 ml) was treated with trimethylsilyl iodide (0.72ml, 5.06 mmoles) and stirred at room temperature under argon for 1.75hours. The mixture was evaporated to dryness and the syrup obtained waspartitioned between ethyl ether (50 ml) and 0.2N hydrochloric acid (2×25ml). The aqueous phase was brought to pH 10 with saturated sodiumbicarbonate (25 ml), treated with solid sodium chloride (2.0 g) andextracted with methylene chloride (3×75 ml) to give 219 mg of titleproduct as a syrup. A second treatment of the aqueous phase with sodiumchloride (2.0 g) and reextraction with methylene chloride (2×100 ml)gave an additional 37 mg of title product. TLC (methylenechloride:methanol, 9:1) R_(f) =0.23.

g)[4S-[4α(R*)7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo[1,4]oxazino-[3,4-b][1,3]thiazepine-7-carboxylicacid, methyl ester

(S)-2-(Acetylthio)benzenepropanoic acid, dicyclohexylamine salt (516 mg,1.32 mmol, 1.2 eq.) was suspended in ethyl acetate (42 ml), washed withpotassium bisulfate (5×6.0 ml) and brine (6.0 ml), dried (anhydrousmagnesium sulfate), filtered, evaporated to dryness and dried in vacuo.

The free acid was dissolved in dry methylene chloride (9.5 ml), cooledto 0° C. (ice-salt bath) and treated sequentially with a solution of theproduct from part (f) (285.5 mg, 1.09 mmol) in dry methylene chloride(4.2 ml), triethylamine (0.14 ml, 1.15 mmol) andbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(484 mg, 1.09 mmol). The reaction mixture was stirred at roomtemperature for 1.0 hour and at room temperature for 2.0 hours underargon, then stripped to dryness. The residual syrup was dissolved inethyl acetate (40 ml), washed with 0.5N hydrochloric acid (2×6.6 ml),water (6.6 ml) and brine (6.6 ml), dried (anhydrous sodium sulfate),filtered, evaporated to dryness and dried in vacuo. The crude productwas chromatographed on silica gel column (Merck), eluting with ethylacetate:hexane mixtures (1:2; 1:1) to give 382.9 mg of title product asa syrup. TLC (ethyl acetate:hexane, 1:1) R_(f) =0.28.

h)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo]1,4]oxazino[3,4-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (g) (382.9 mg, 0.82 mmol) inmethanol (9.0 ml) was purged with argon for 30 minutes, cooled to 0° C.(ice-salt bath) then treated with 1.0N sodium hydroxide (3.32 ml, 4.0eq; previously purged with argon for 30 minutes), maintaining thebubbling of argon throughout the addition and length of the reaction.The reaction mixture was stirred at 0° C. for 5.0 hours and at roomtemperature for 1.0 hour, brought to pH 2.0 with 5% potassium bisulfate(14.5 ml), warmed to room temperature and extracted with ethyl acetate(2×50 ml). The combined organic extracts were washed with brine (10 ml),dried (anhydrous sodium sulfate), filtered, evaporated to dryness anddried in vacuo. The syrup obtained was evaporated twice from hexane (25ml) and the solid foam obtained was chromatographed on a silica gelcolumn (Merck), eluting the column with toluene:acetic acid mixtures(100:1; 50:1; 20:1) to give 212 mg of title product as a solid; m.p.224°-226° C.; [α]_(D) =-50° (c=0.45, methanol). TLC (toluene:aceticacid, 5:1) R_(f) =0.28. HPLC: t_(R) =5.37 min. (UV 220 nm) (98.9%); YMSS-3 ODS (C-18) 6×150 mm; 60% (10% water-90% methanol-0.2% phosphoricacid)/40% (90% water-10% methanol-0.2% phosphoric acid), isocratic.

Anal. calc'd for C₁₈ H₂₂ N₂ O₅ S₂ •0.14 H₂ O: C, 52.34; H, 5.44; N,6.78; S, 15.53 Found: C, 52.58; H, 5.57; N, 6.44; S, 15.16.

EXAMPLE 22[4S-[4α(R*),7α10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid

The product of Examples 3 and 11 was also prepared as follows:

a)[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester, p-toluenesulfonic acid salt

[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester (6.11 g) was dissolved in ethyl acetate (about 100ml) and treated with a solution of p-toluenesulfonic acid monohydrate(4.52 g) in methanol (3 ml) and ethyl acetate (20 ml). A precipitateformed immediately. The mixture was diluted with additional ethylacetate and the solid was collected by filtration. The solid was washedwith ethyl ether and dried in vacuo to give 7.908 g of the title productas a pale yellow solid in 98% purity; m.p. 179°-181° C. (decomp.).

b)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido-[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A slurry of the product from part (a) (636 mg, 1.48 mmol) in methylenechloride (5 ml) and dimethylformamide (1 ml) was treated withN-methylmorpholine (163 μl, 150 mg, 1.48 mmol) followed by1-hydroxy-7-azabenzotriazole (208 mg, 1.52 mmol). The bright yellowsolution was then treated with (S)-2-(acetylthio)benzenepropanoic acid(333 mg, 1.48 mmol) in methylene chloride (5 ml) and cooled in anice-bath. Ethyl-3-(dimethylamino)propyl carbodiimide, hydrochloride salt(2.88 mg, 1.50 mmol) was added and the mixture was stirred at 0° C. for1 hour and at room temperature for 1.5 hours. The solvent was removed byrotary evaporation and the residue was partitioned between ethyl acetateand 0.5N hydrochloric acid. The ethyl acetate extract was washedsuccessively with water (twice), 50% saturated sodium bicarbonate, andbrine, then dried (sodium sulfate), filtered, and stripped to give 651.2mg of title product as a white foam.

c)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid

A solution of the methyl ester product from part (b) in deoxygenatedmethanol was treated with 1N sodium hydroxide according to the procedureof Example 3(d) to afford the title product.

EXAMPLE 23[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid

The product of Examples 3, 11 and 22 was also prepared as follows:

a) N-[(Phenylmethoxy)carbonyl]-L-methionine

In a 2 L flask equipped with a mechanical stirrer and internalthermometer, sodium hydroxide (61.65 g, 1.541 mol) was dissolved indistilled water (1000 ml). To this solution, L-methionine (100.0 g,0.670 mol) was added at room temperature. The solution was cooled in anice bath (internal temperature 3° C.) and benzyl chloroformate (110 ml,0.737 mol) was added over 10 minutes. After a 15 minute inductionperiod, the internal temperature rose from 3° C. to 12° C. over 30minutes and then dropped to 0° C. over 15 minutes. The reaction wasstirred at 0° C. for 2 hours, during which time the initially cloudyreaction mixture became homogeneous. The ice bath was removed, and thereaction was allowed to warm to room temperature over 1 hour. Thereaction mixture was transferred into a separatory funnel and washedwith hexane (2×300 ml). The aqueous layer was acidified with 6Nhydrochloric acid to pH 5 and diluted with ethyl acetate (600 ml). Themixture was further acidified to pH 2. The organic layer was separated,and the aqueous layer was extracted with ethyl acetate (3×600 ml). Theorganic extracts were combined and washed with brine (750 ml), dried(magnesium sulfate), filtered and concentrated in vacuo to produce alight yellow oil. The crude product (oil) was dissolved in toluene (1500ml), and the solution was concentrated to half of its volume. A secondportion of toluene (750 ml) was added, and concentrated again such that630 ml of toluene remained. This solution was stored at 5° C. overnight,during which time some of the product crystallized from solution. Thesolid was redissolved by warming to room temperature. Toluene (134 ml)was then added (a few seed crystals remained). With mechanical stirring,heptane (500 ml) was added in 30 ml portions at 10 minute intervals(approximately 3 hours total addition time). At this point, the productstarted to crystallize from solution. An additional portion of heptane(1020 ml) was added over 1.5 hours, and the resulting slurry was stirredfor 2 hours. The product was collected by vacuum filtration, washed with1:2 toluene:heptane (3×150 ml) and heptane (3×500 ml) and air-dried togive 158.6 g of title product as a white solid; m.p. 66° C.; [α]_(D)=-1.5° (c=1, 95% ethanol). TLC (ethanol:water, 3:1) R_(f) =0.78.

Anal. calc'd for C₁₃ H₁₇ NO₄ S: C, 55.11; H, 6.05; N, 4.94 Found: C,54.96; H, 6.20; N, 4.83.

b) N-[(Phenylmethoxy)carbonyl]-L-methionine, methyl ester

In a 3 L flask equipped with a mechanical stirrer and an argon inlet,the product from part (a) (100.0 g, 0.353 mol) was dissolved in methanol(2 L), and p-toluenesulfonic acid monohydrate (6.71 g, 0.035 mol) wasadded. The reaction mixture was stirred under argon for 21 hours.Triethylamine (4.9 ml, 0.035 mol) was added, and the reaction mixturewas stirred for an additional 15 minutes. The reaction mixture wasconcentrated in vacuo to a pale yellow oil. The oil was dissolved inethyl acetate (900 ml), and the solution was washed with 1N hydrochloricacid (740 ml), saturated sodium bicarbonate (2×740 ml) and brine (740ml). The organic layer was dried (magnesium sulfate), filtered andconcentrated in vacuo to a light yellow oil. The oil was concentratedfrom hexane (2×100 ml) to obtain 98.22 g of title product as a whitesolid.

c) N-[(Phenylmethoxy)carbonyl]-L-methionine, sulfoxide, methyl ester

In a 3 L flask equipped with mechanical stirrer, the product from part(b) (97.95 g, 0.329 mol) was dissolved in methanol (1675 ml) anddistilled water (215 ml). The solution was cooled in an ice bath, andsodium bicarbonate (28.5 g, 0.339 mol) was added. N-Chlorosuccinimide(44.0 g, 0.329 mol) was added in small portions over 25 minutes so thatthe internal temperature did not exceed 7° C. The mixture was stirred inan ice bath for 1 hour and then was allowed to warm to room temperatureover 1 hour. The mixture was concentrated in vacuo by about 75% toremove the methanol, diluted with ethyl acetate (1000 ml) and washedwith brine (500 ml). The brine layer was back-extracted with ethylacetate (2×200 ml). The organic extracts were combined, dried (magnesiumsulfate), filtered, and concentrated in vacuo to a clear, viscous oil.The oil was concentrated from toluene (3×100 ml) and residual solventswere removed under high vacuum to produce crude title product as a clearoil, which solidified to a white solid (131.4 g). The crude productcontained 12 weight percent succinimide and 9 weight percent toluene byNMR. The product was a mixture of sulfoxide diastereomers.

d) S-[(Acetyloxy)methyl]-N-[(phenylmethoxy)carbonyl]-L-homocysteine,methyl ester

To a 1 liter flask containing the product from part (c) (102.8 gcorrected weight, 0.328 mol) was added toluene (480 ml), sodium acetate(32.3 g, 0.394 tool) and acetic anhydride (186 ml, 1.970 tool). Theresulting mixture was refluxed (118° C.) under argon for 18 hours. Thedark brown reaction mixture was allowed to cool to room temperature.After an hour at room temperature the reaction mixture became very thickwith solids. The solids were dissolved with ethyl acetate (100 ml), andthe mixture was partially concentrated in vacuo to a viscous brownresidue. The residue was concentrated from toluene (240 ml) to removeacetic anhydride, diluted with ethyl acetate (1000 ml), and carefullywashed with saturated sodium bicarbonate (4×680 ml). The organic layerwas washed with brine (450 ml), dried (magnesium sulfate), filtered andconcentrated in vacuo. Residual solvents were removed under high vacuumto produce a light brown solid. The crude product was dissolved inn-butyl acetate (450 ml) with warming (35° C.) and stirring. Aftercooling to room temperature, hexane (200 ml) was added slowly to thesolution with stirring over 15 minutes. At this point the productcrystallized from the solution. An additional portion of hexane (700 ml)was added over 30 minutes, and the resulting slurry was stirred for 3hours. The product was collected by filtration and washed with 1:2n-butyl acetate:hexane (200 ml), 1:4 n-butyl acetate:hexane (2×240 ml),and hexane (2×250 ml). The product was air-dried, then dried under highvacuum to give 87.7 g of title product as a pale brown solid; m.p. 73°C.; [α]_(D) =-1.6° (c=1, 95% ethanol). TLC (5% methanol/methylenechloride) R_(f) =0.80.

Anal. calc'd for C₁₆ H₂₁ NO₆ S: C, 54.07; H, 5.95; N, 3.94 Found: C,53.48; H, 5.74; N, 3.82.

e) S-Acetyl-N-[(phenylmethoxy)carbonyl]-L-homocysteine

In a 1 L flask, a solution of the product from part (d) (83.0 g, 0.233mol) in tetrahydrofuran (415 ml) was sparged with argon for 30 minutes.In a separate 2 L flasked equipped with a mechanical stirrer and anargon inlet, a solution of 86.8% potassium hydroxide (62.7 g, 0.969 mol)in distilled water (280 ml) was sparged with argon for 15 minutes.

The tetrahydrofuran solution was added to the potassium hydroxidesolution (internal temperature 20° C.) rapidly, via cannula, withvigorous stirring under argon. The flask containing the product frompart (d) was rinsed with 20 ml of tetrahydrofuran (sparged with argonfor 15 minutes) and the rinse was added to the reaction mixture. After30 minutes, the reaction was clear and biphasic, and an exotherm to 28°C. had occurred.

After an additional 2 hours, the reaction was cooled to l° C. (internal)and sodium borohydride (2.75 g, 0.073 mol) was added in one portion(exotherm to 6.8° C.). The reaction mixture was stirred for anadditional 20 minutes at 0° C. and then allowed to warm to 11° C. over30 minutes. The reaction mixture was cooled to 1° C., and aceticanhydride (68.6 ml, 0.727 mol) was added over 10 minutes. An exotherm to10° C. occurred during the addition. The internal temperature droppedback to 4° C. before the addition was complete. The cooling bath wasremoved, and the reaction was stirred at ambient temperature for 45minutes.

The reaction mixture was concentrated in vacuo to approximately half ofits volume, acidified to pH 2 with 6N hydrochloric acid (175 ml), andextracted with ethyl acetate (2×1.1 L). The combined organic extractswere washed with brine (560 ml). The organic layer was treated withactivated carbon and anhydrous magnesium sulfate, filtered, andconcentrated in vacuo to a yellow oil. n-Butyl acetate (380 ml) wasadded, and the solution was concentrated in vacuo (45° C.) to half ofits volume. A second portion of n-butyl acetate (190 ml) was added andconcentrated again such that 190 ml of n-butyl acetate remained. Heptane(300 ml) was added slowly with stirring to haziness, and seed crystalswere added. After 15 minutes a white solid crystallized from thesolution. A second portion of heptane (570 ml) was added slowly over 30minutes, and the resulting slurry was stirred at room temperatureovernight. The product was collected by filtration, washed with 1:3n-butyl acetate:heptane (2×275 ml) and hexane (2×275 ml), air-dried, andthen dried under high vacuum to produce 50.1 g of title product as awhite solid; m.p. 73°-74° C.; [α]_(D) =-1.3° (c=1, 95% ethanol). TLC(ethanol:water, 3:1) R_(f) =0.83.

Anal. calc'd for C₁₄ H₁₇ NO₅ S: C, 54.01; H, 5.50; N, 4.50 Found: C,53.88; H, 5.45; N, 4.44.

The filtrate was concentrated so that 100 ml of butyl acetate remained.This solution was treated with 310 ml of heptane as described above toobtain a second crop of 8.4 g of title product as a white solid for atotal yield of 58.5 g.

f)[S-(R*,R*)-2-(Acetylthio)-1-oxo-2-[[(phenylmethoxy)carbonyl]-[[4-(phenylmethoxy)carbonyl]amino]butyl]amino]-6,6-dimethoxyhexanoicacid, methyl ester

S-Acetyl-N-[(phenylmethoxy)carbonyl]-L-homocysteine (0.456 mol) wasdissolved in a mixture of methylene chloride (600 ml) anddimethylformamide (90 ml), and hydroxybenzotriazole hydrate (64.72 g,0.479 mol) was added. The mixture was cooled in an ice-bath and asolution of (S)-2-amino-6,6-dimethoxyhexanoic acid, methyl ester[prepared as described in Example 1(e), 93.7 g, 0.456 mol) dissolved inmethylene chloride (600 ml) was added. Finally,ethyl-3-(dimethylamino)propylcarbodiimide, hydrochloride salt (91.83 g,0.479 mol) was added and the reaction was stirred for one hour at 0° C.,then for 2 hours at room temperature. At the end of that time, thereaction mixture was concentrated in vacuo and the residue waspartitioned between ethyl acetate (3 L) and saturated aqueous sodiumbicarbonate (1 L). The organic extract was washed with water (1 L), 5%potassium bisulfate (1 L), water (1 L), and brine (1 L), then dried(sodium sulfate) and concentrated in vacuo to 238 g of crude product.The crude product was dissolved in ethyl acetate:methylene chloride(1:1, 300 ml) and applied to a 10×15 cm pad of Merck silica gel. Elutionwith 8:2 ethyl acetate:hexane (7 L) followed by ethyl acetate (4 L)provided 205.28 g of title product.

g)[4S-(4α,7α,10aβ)]-Octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid methyl ester

A solution of the product from part (f) [205.28 g, 0.412 mol, dried byevaporating in methylene chloride/toluene) in methanol (2 L) was cooledto 0° C. (ice bath) and purged with argon for 30 minutes. A 25% byweight solution of sodium methoxide in methanol (95.1 ml, 1.01 eq.) wasadded rapidly with continued argon purging, and the reaction was stirredfor 10 minutes longer, then quenched by the addition of 1 L of saturatedammonium chloride solution, diluted with 0.5 L of water, and treatedwith 3 L of ethyl acetate. The resulting mixture was divided into twoportions which were each separately concentrated in vacuo to removeorganics (ethyl acetate and methanol). The concentrated residues wererecombined and treated with 1 L of ethyl acetate. The organic layer wasseparated and rinsed with 0.5 L of saturated ammonium chloride. Thecombined aqueous solutions were reextracted with 1 L of ethyl acetate.The organic extracts were combined and washed with 1 L of water and two1 L portions of brine, dried (sodium sulfate), filtered andconcentrated. The residue was further evaporated with methylene chlorideand dried in vacuo to give 182.65 g of free sulfhydryl of the productfrom part (f).

This free sulfhydryl intermediate (0.400 mol) was dissolved in methylenechloride (4 L) and treated with 30.8 ml (0.400 mol) of trifluoroaceticacid. The reaction mixture was refluxed for 16 hours, then cooled andconcentrated in vacuo. The resulting residue was dissolved in 2 L ofethyl acetate, then washed with 400 ml of 0.1N hydrochloric acid, 1 L ofwater, 1 L of saturated sodium bicarbonate, 1 L of water, and 1 L ofbrine, dried (sodium sulfate), filtered and concentrated. The residuewas evaporated with methylene chloride and dried in vacuo to afford166.24 g of title product.

h) [4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7-H-pyrido[2,1-b][1,3]thiazepine-7-carboxylic acid, methyl ester

Iodotrimethylsilane (76.6 ml, 0.538 mol) was added to a solution underargon containing the product from part (g) (162.43 g, 0.414 mol)dissolved in methylene chloride (1.5 L). After stirring for 1.5 hours,the reaction mixture was concentrated in vacuo and the residue waspartitioned between 1 L of ethyl acetate and 700 ml of 1N hydrochloricacid (evolution of CO₂ occurs; pH 1.2). The ethyl acetate layer wasseparated and extracted with 300 ml of 1N hydrochloric acid. Thecombined acidic aqueous extracts were washed with a further 1 L of ethylacetate, then cooled to 0° C. and basified with 4N sodium hydroxide(about 275 ml) to pH 10.0. The aqueous layer was saturated with solidsodium chloride, then extracted with five 1 L portions of methylenechloride. The combined organic extracts were dried (sodium sulfate),filtered and concentrated in vacuo. The residue was redissolved in 1 Lof methylene chloride and rinsed with 0.5 L of brine, dried (sodiumsulfate), filtered and concentrated to give 98.8 g of title product.

i)[4S-[4α(R*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido-[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

(S)-2-(Acetylthio)benzenepropanoic acid, dicyclohexylamine salt (173.1g, 0.427 mol) was partitioned between ethyl acetate (1 L) and 10%potassium bisulfate (800 ml). The organic layer was separated and washedwith 5% potassium bisulfate (1 L), 50% brine (1 L) and brine (1 L),dried (sodium sulfate), filtered and concentrated in vacuo. The residuewas evaporated several times with methylene chloride, then driedovernight in vacuo to yield 97.3 g of crude(S)-2-(acetylthio)benzenepropanoic acid.

A solution of this (S)-2-(acetylthio)benzenepropanoic acid (0,427 mol)dissolved in methylene chloride (900 ml) was cooled in an ice-bath andtreated with a solution of the product from part (h) (100.28 g, 0.388mol) in methylene chloride (600 ml), triethylamine (154.1 ml, 0.388mol), and finally benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (188.9 g, 0,427 mol) added in one portion. After onehour at 0° C. and 2 hours at room temperature, the reaction mixture wasconcentrated in vacuo and dissolved in 2 L of ethyl acetate. The organicsolution was concentrated in vacuo and dissolved in 2 L of ethylacetate. The organic solution was washed with 0.5 L of brine, 1 L of0.5N hydrochloric acid, 1 L of water, 2 L of saturated sodiumbicarbonate, 1 L of water, and 1 L of brine, dried (sodium sulfate),filtered and concentrated. At this point, those aqueous rinses whichcontained product (TLC indication) were reextracted with ethyl acetate.The ethyl acetate extracts were worked up in the usual manner and allcombined to give a crude yellow oil product. The yellow oil was appliedto a 15×15 cm silica gel pad prepared in 1:1 ethyl acetate:hexanes andeluted with 7 L of 1:1 ethyl acetate:hexanes followed by 4 L of 6:4ethyl acetate:hexanes and finally 2 L of 7:3 ethyl acetate:hexanes. Thefiltrates containing the desired product were concentrated to give123.57 g of title product.

j)[4S-4α(R*),7α,10aβ]]-Octahydro-4-[(2-mercapto-1-oxo-3-phenypropyl)amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid

Into a 12 l three-necked flask, fitted with an additional funnel andmechanical stirrer, was placed a solution of the product from part (i)(96.0 g, 0.207 mol) in methanol (1.1 l). The solution was purged withargon for 30 minutes then cooled in an ice-bath until the internaltemperature was +7° C. A total of 1.45 L of 1N sodium hydroxide solution(previously sparged with argon for 30 minutes) was added over 1 hour.The reaction mixture was continuously sparged with argon during theaddition. The reaction temperature rose to +12° C. and was maintainedduring the addition. The reaction mixture was stirred for an additional30 minutes, then warmed to room temperature with an ambient water bathand stirred with sparging for 2.5 hours. About 250 ml of 6N hydrochloricacid was added dropwise over 15-20 minutes to adjust the pH to 2. Agummy precipitate formed during the acidification. After continualstirring for a further 2 hours, the precipitate changed to a fine whitesolid, with the presence of some larger chunks of solid product. Theproduct was collected on a 600 ml sintered glass funnel. Washing thecollected solid with 1 L of water followed by 2 L of anhydrous ether andfinal drying in vacuo afforded 70.3 g of title product as a fine whitesolid; m.p. 218°-220° C. (dec.). TLC (1:99 acetic acid/ethyl acetate)R_(f) =0.48. HPLC: t_(R) (YMC S-3 ODS 6.0×150 mm; 1.5 mL/min., isocratic60% B, Buffer A=methanol/water/phosphoric acid (10:90:0.2), BufferB=methanol/water/phosphoric acid (90:10:0.2))=9.33 min., 99.3% of totalpeak area at 220 nm.

Anal. calc'd for C₁₉ H₂₄ N₂ O₄ S₂ : C, 55.86; H, 5.92; N, 6.86; S, 15.70Found: C, 55.83; H, 5.83; N, 6.96; S, 15.70.

EXAMPLE 24[4S-[4α(R*),7α10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid, methyl ester

The coupling reaction described in Examples 3(c), 11(i), 22(b), and23(i) was also carried out as follows:

A solution of (S)-2-(acetylthio)benzenepropanoic acid (1.83 g, 8.14mmol) and[4S-(4α,7α,10aβ)]octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester (2.11 g, 8.17 mmol) in dry methylene chloride (20 ml)was cooled to 0° C. and ethyl-3-(dimethylamino)propyl carbodiimide,hydrochloride salt (1.77 g, 9.32 mmol) was added in a single portion.The reaction mixture was stirred at 0° C. for 6 hours and then wasconcentrated to an oily foam. The residue was then partitioned betweenethyl acetate (100 ml) and 1N hydrochloric acid (50 ml). The organicphase was washed with 1N hydrochloric acid (50 ml), saturated aqueoussodium bicarbonate (2×50 ml), and saturated aqueous sodium chloride (50ml), dried (anhydrous sodium sulfate), filtered and concentrated invacuo to give 3.43 g of title product as a white foam.

EXAMPLE 25[4S-(4α(R*),7α,10aβ)]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, 1,1-dimethylethylamine salt

A 15 ml, 3-necked flask equipped with a reflux condenser was evacuatedand refilled with argon three times.[4S-[4α(R*),7α,10aβ]-Octahydro-4-[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid (0.20 g) and a 1:1 solution of degassed absolute ethanol andacetonitrile (1.0 ml) were charged into the flask. As the heterogeneousmixture was stirred, tert-butylamine (53.0 μl, 1.03 eq.) was addeddropwise. The solution became homogeneous within three minutes after theamine addition was completed. The solution (internal temperature of 20°C.) was diluted slowly by the dropwise addition of acetonitrile to afinal volume of 10 ml. After an additional 2 hours of stirring, thesolids were filtered, washed once with 100% acetonitrile (5 ml), airdried, and placed under high vacuum for 2 hours to remove residualsolvents to give 0.2 g of title product as a white crystalline solid.

The above material was combined with material from other runs andrecrystallized as follows. A 25 ml, 3-necked flask equipped with areflux condenser, magnetic stirrer bar, and addition funnel wasevacuated and refilled with argon three times. The batches of1,1-dimethylamine salt product (0.37 g) and 59% acetonitrile/ethanol(2.28 ml) were added to the flask. The flask and contents were warmed to29°-32° C. to dissolve the solids. The solution was diluted withacetonitrile (27 ml). The heating bath was removed, and the flask wasallowed to cool to room temperature (20° C.). After one hour ofadditional stirring, the mixture was filtered, and the solids werewashed once with acetontrile (10 ml), air-dried, and placed under highvacuum to remove residual solvents and give 0.29 g of title product as awhite crystalline solid; m.p. shrinks at 160° C. and slowly melts anddecomposes as the temperature is increased to 190°-191° C. (at 190°-191°C., the remaining glossy material melts rapidly).

EXAMPLE 26[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

This intermediate of Examples 3(c), 11(h) and 23(h) was also prepared asfollows:

a) 2-(Acetylamino)-2-[4-(acetyloxy)butyl]propanedioic acid, diethylester

A stirred suspension of 95% sodium hydride (60.8 g, 2.532 mol) inanhydrous dimethylformamide (500 ml) under an atmosphere of argon wascooled to 0° C. (ice bath). A solution of diethyl acetamidomalonate (500g, 2.302 mol) in anhydrous dimethylformamide (1.2 l) was added over aperiod of 45 minutes while keeping the reaction temperature below 18° C.After the addition was complete, the turbid solution was graduallywarmed to room temperature. After stirring for one hour at roomtemperature, 4-bromobutyl acetate (471.5 g, 2.417 mol) was added. Themixture was then stirred at 59°-60° C. for 18 hours. The resultingslurry was cooled to room temperature, quenched with absolute ethanol(40 ml) and glacial acetic acid (4 ml), stirred for about 15 minutes,poured into a 10% lithium chloride solution and extracted with ethylacetate (2×3 l). The combined ethyl acetate extracts were washed with10% lithium chloride (3×3 l), dried (anhydrous sodium sulfate), andevaporated in vacuo to give 750 g of title product as an oil.

b) 2-(Acetylamino)-6-hydroxyhexanoic acid

The product from part (a) (730 g, 2.2 mol) was weighed into a 5 l.3-neck flask (equipped with a thermometer, magnetic stirrer and aircooled condenser) and diluted with absolute ethanol (300 ml) followed bythe addition of aqueous 6N sodium hydroxide (1.6 l, 9.6 mol). Thereaction mixture was heated at 68°-70° C. for 5 hours and a homogenoussolution was obtained. The reaction was cooled to room temperature, and6N hydrochloric acid (1.32 l) was added slowly to pH 1.3. The flask wasequipped with a short path still head to distill off the ethanol as thetemperature was slowly increased to 87°-90° C. and maintained at thistemperature for 8.5 hours. Slow carbon dioxide evolution was observed.The total volume of distillate was 600 ml. The pH of the final solutionwas 3.0. The reaction mixture was concentrated in vacuo until all of thewater evaporated off and then concentrated from toluene (2×500 ml). Thesemi-solid mass was triturated with absolute ethanol (1 l), filtered,and rinsed with additional absolute ethanol (500 ml). The filtrate wasconcentrated in vacuo to yield 509 g of crude oil (82% purity) whichcontained ethanol and toluene.

c) (S)-2-Amino-6-hydroxyhexanoic acid

The crude product from part (b) (443 g, includes some toluene andethanol, starting material weight estimated to be 394 g) was dissolvedin water (3.3 l) and 1N lithium hydroxide was added until the pH was 7.5(1.53 l required). The mixture was heated to 35° C. and acylase (grade 1from porcine kidney, 0.4 g) was added and the reaction mixture wasstirred for 24 hours. At the end of this time period the pH was 7.33.The pH was readjusted to 7.5 with 1N lithium hydroxide (about 2 ml),additional acylase (0.4 g) was added, and the reaction was stirred for17 more hours (pH 7.3). The pH of the solution was adjusted to 5.9 withacetic acid. Celite® (20 g) and charcoal (20 g) were added and thereaction was heated to 92° C. and maintained for 5 minutes. The reactionwas filtered through a pad of Celite® and concentrated in vacuo to asemi-paste (441 g). This was triturated with 900 ml of 1:5:10water:ethanol:dimethylformamide. Some warming was required to break upthe original cake. The reaction mixture was refrigerated overnight,filtered, and washed with 200 ml. of the above solvent mixture to yield214 g of crude material (about 40% N-acetyl material). This material wassuspended in methanol (500 ml), warmed on a steam bath, allowed to standfor 2 hours, and filtered. This procedure was repeated a second time toyield 108 g of title product; [α]_(D) =+22° (c=1.44, 6N hydrochloricacid).

Alternatively, steps (b) and (c) were also performed as follows:

b) 2-(Acetylamino)-6-hydroxyhexanoic acid

A 5 l, 3-necked flask, equipped with a mechanical stirrer andthermocouple thermometer, was charged with the product from part (a)(631 g, 1.933 mol) and tetrahydrofuran (259 ml). A 6N sodium hydroxidesolution (1385 ml, 8.31 mol) was added to the stirred solution over 40minutes. A strong exotherm occurred and it was necessary to cool thereaction mixture in an ice bath to keep the temperature under 60° c. Thereaction mixture was then heated to slight reflux (pot temperature at67°-68° C.) for 5.5 hours.

The mixture was stirred at room temperature overnight (16.5 hours). ThepH was brought from 12.75 to 1.30 with the gradual addition of 6Nhydrochloric acid solution (1150 ml, 6.9 mol), maintaining thetemperature at about 25° C. The mixture was heated gradually with ashort distillation head until distillation and gas (carbon dioxide)evolution started (72.3° C. pot temperature, 70° C. head temperature)and until distillation stopped and gas evolution became very slow (94.1°C. pot temperature, 50° C. head temperature). Total distillate collectedwas 410 ml and the pot residue had a pH of 3.9. Heating was continuedfor another ten minutes with no additional gas evolution. Total heatingtime from the start of distillation was 7.5 hours.

After stirring at room temperature overnight, the clear reaction mixture(pH 3.50) was stripped in the rotary evaporator under vacuum in a 60° C.bath and the pasty residue was stirred with absolute ethanol (750 ml).The resulting crystalline suspension was stripped in the rotaryevaporator (pump vacuum, 60° C. bath) and the pasty residue was chasedwith absolute ethanol (2×750 ml). To the final residue, absolute ethanol(1500 ml) was added and the mixture was stirred in a 60° C. bath untilit became a fine crystalline suspension, about 20 minutes, and thenstirred at room temperature for 20 minutes. The suspension was filteredand the cake was washed with absolute ethanol (2×300 ml). The filtratesappeared hazy and were further clarified by filtration through a pad ofCelite®. The new, clear filtrate was stripped in the rotary evaporatorto give 434.6 g of title product as an amber-colored thick syrup. TLC(10:1:1, methanol:acetic acid:water) R_(f) =0.59.

c) (S)-2-Amino-6-hydroxyhexanoic acid

A 5 l, 3-necked flask, equipped with a mechanical stirrer andthermometer, was charged with the product from part (b) (434 g ,1.93mol), and water (3 l). The pH of the hazy solution was adjusted from4.05 to 7.50 by the addition of 1N lithium hydroxide (705 ml). Thesolution was warmed to 36° C. and porcine kidney acylase I (0.710 g) wasadded. The mixture was stirred at 35° to 36° C. for 23.5 hours, Thereaction mixture was cooled to room temperature and the pH was broughtfrom 7.0 to 5.9 by the addition of glacial acetic acid (4.4 ml). Celite®(29 g) and charcoal (29 g) were added and the temperature was raisedwith stirring to 91° C. The heating was removed and the mixture wasallowed to cool to room temperature. The suspension was filtered throughan 18.5 cm filter paper disc and the cake was thoroughly washed withwater. The colorless flitrates (about 3.9 l) were concentrated in arotary evaporator at 60° C. to give 476 g of a clear, thick oil.Absolute ethanol (720 ml) was added and the mixture was stirred until itbecame a homogeneous crystalline suspension. The solvent was againstripped off and absolute ethanol (1584 ml) was added to the white solidresidue. The suspension was rolled in the rotary evaporator at roomtemperature overnight (15 hours) and filtered through 18.5 cm paper. Thecake was washed with absolute ethanol (7×100 ml) and dried to constantweight under vacuum to give 85.8 g of white, crystalline title product.TLC (methanol:water:acetic acid, 10:1:1) R_(f) =0.62.

Anal. calc'd for C₆ H₁₃ NO₃ : C, 48.25; H, 8.94; N, 9.38 Found: C,48.66; H, 8.77; N, 9.43.

d)[S-(R*,R*,)-2-[[4-(Acetylthio)-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6-hydroxyhexanoicacid, methyl ester

A slurry of (S)-2-amino-6-hydroxyhexanoic acid (1.0 g, 6.8 mmol) inmethanol (20 ml) was stirred under argon at room temperature and treatedwith trimethylsilyl chloride (1.9 ml, 15 mmol). The resulting solutionwas stirred at room temperature for 18 hours. The solvent volume wasreduced to about 3.5 ml under reduced pressure. Acetonitrile (5 ml) wasadded and the solution was cooled to -10° C. N,N-Diisopropylethylamine(4.15 ml, 23.8 mmol) was then added and the solution was cooled to -40°C. to give a solution containing (S)-2-amino-6-hydroxyhexanoic acid,methyl ester.

In a separate flask, a solution ofS-acetyl-N-[(phenylmethoxy)carbonyl]-L-homocysteine [prepared asdescribed in Example 23(e), 2.117 g, 6.8 mmol] in acetonitrile (5 ml) at0° C. was treated with N,N-diisopropylethylamine (1.20 ml, 6.8 mmol). Inanother flask, hydroxybenzotriazole hydrate (0.104 g, 0.68 mmol) andmethanesulfonyloxybenzotriazole (1.450 g, 6.8 mmol) were dissolved inacetonitrile and cooled to -18° C. The previously formedS-acetyl-N-[(phenylmethoxy)carbonyl]-L-homocysteine was then addeddropwise to this solution while maintaining the internal temperature atless than -10° C. After stirring at -18° to -12° C. for three hours, theresulting solution was added dropwise to the above solution containing(S)-2-amino-6-hydroxyhexanoic acid, methyl ester at -40° C. The mixturewas allowed to slowly warm to 16° C. over 18 hours. The reaction wasthen poured into ethyl acetate (50 ml) and 1N hydrochloric acid (50 ml).The mixture was transferred into a separatory funnel and the layers wereseparated. The aqueous layer was extracted with ethyl acetate (2×50 ml).The organic layers were combined and then washed with iN hydrochloricacid (100 ml), saturated sodium bicarbonate (100 ml), and saturatedsodium chloride (100 ml). The solution was dried over magnesium sulfate,filtered, and concentrated to a white solid. To this solid was addedtert-butyl methyl ether (20 ml) and the resulting slurry was stirred atroom temperature for 4 hours and filtered. The product was washed withtert-butyl methyl ether and dried to yield 2.087 g of title product;m.p. 89 °-90° C.

e)[[S-(R*,R*)]-2-[[4-(Acetylthio)-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6-oxohexanoicacid, methyl ester

To a solution of oxalyl chloride (546 μl, 6.27 mmol) in dry methylenechloride (16 ml) at -65° C. (internal temperature) was added dropwise asolution of dimethylsulfoxide (905 μl, 12.54 mmol) in methylene chloride(13 ml) over 12 minutes while maintaining an internal temperaturebetween -65° and -60° C. A solution of the product from part (d) (1.90g, 4.18 mmol) in methylene chloride (7 ml) was added to the reactionflask over 20 minutes producing a turbid mixture. Additional methylenechloride (1 ml) was used to complete the transfer of the alcohol intothe reaction flask and the reaction was allowed to stir at -65° C. for40 minutes. Next N,N-diisopropylethylamine (3.7 ml, 20.90 mmol) wasadded thus producing a clear solution. After stirring an additional 30minutes at -65° C., the reaction was allowed to warm to -18° C. over 2hours. The reaction was quenched with 10% aqueous potassium bisulfate(30 ml) and then warmed to room temperature. The reaction mixture wasdiluted with 25 ml of water, mixed, and the phases were separated. Theaqueous fraction was back-extracted with methylene chloride (2×25 ml).The combined organic extracts were washed with 10% aqueous potassiumbisulfate (25 ml), saturated aqueous sodium bicarbonate (2×25 ml), brine(25 ml), dried (magnesium sulfate), filtered and concentrated in vacuoto give 1.84 g of title product as a white solid.

f)[4S-(4α,7α,10aβ)]-Octahydro-4-[[(phenylmethoxy)carbonyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A dried flask under argon was charged with the product from part (e)(1.76 g, 3.89 mmol) and methanol (17 ml). The solution was cooled to 0°C. and sparged with argon for 25 minutes. Sodium methoxide solution (25%by weight in methanol, 983 μl, 4.28 mmol) was added to the reactionmixture over about 15 seconds. The reaction was quenched after one hourwith 1N hydrochloric acid solution (20 ml) and then allowed to warm toroom temperature. Ethyl acetate (35 ml) was added and after mixing, thelayers were separated. The aqueous fraction was back-extracted withethyl acetate (2×15 ml). The combined organic fractions were washed with1N hydrochloric acid solution (15 ml), brine, dried (magnesium sulfate),filtered, and concentrated in vacuo to give 1.69 g of[S-(R*,R*)]-2-[[4-mercapto-1-oxo-2-[[(phenylmethoxy)carbonyl]amino]butyl]amino]-6-oxohexanoicacid, methyl ester as a white foam.

A solution of this white foam and trifluoroacetic acid (305 μl, 3.95mmol) in methylene chloride (17 ml) was refluxed for 2.25 hours. Aftercooling to room temperature, the reaction was concentrated and theresidue was dissolved in ethyl acetate (25 ml), washed with saturatedsodium bicarbonate solution (2×20 ml) and brine, dried (magnesiumsulfate), filtered and concentrated in vacuo to yield 1.50 g of titleproduct as a white foam.

g)[4S-(4α,7α,10aβ)]-Octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

Treatment of the product from part (f) with iodotrimethylsilaneaccording to the procedure of Example 23(h) or 11(h) removes theN-protecting group and yields the desired 4-amino product.

EXAMPLE 27

(S)-2-Phthalimido-6-hydroxyhexanoic acid

This intermediate of Example 1(c) was also prepared as follows:

To a solution of 2-(acetylamino)-2-[4-(acetyloxy)butyl]propanedioicacid, diethyl ester [730 g, 2.2 mol, prepared as described in Example24(a)] in absolute ethanol (300 ml) was added 6N sodium hydroxidesolution (1.6 l). The reaction was heated at 70°-75° C. for 5 hours,then at 90° to 95° C. to distill off most of the ethanol. The reactionwas cooled and acidified to pH 1.3 using 6N hydrochloric acid (about 1.3l), then heated at 90°-100° C. to achieve decarboxylation. Uponcompletion, the crude reaction mixture was cooled to room temperature.

The above crude reaction mixture was heated to 35° C. and treated withabout 600 ml of 6N sodium hydroxide followed by 1N sodium hydroxide toadjust to pH of 7.5 (final volume was about 5.3 l). To this mixture wasadded 0.6 g of porcine kidney acylase I. After stirring overnight at 35°C. the pH was 7.25. The pH was adjusted to 7.5 and an additional 300 mgof acylase was added. After stirring overnight, the reaction appeared tobe about 90% complete. The reaction mixture was next treated with 20 gof charcoal and 20 g of Celite®, then heated to 85° C. and maintained atthat temperature for 10 minutes, then cooled to 50° C. and filtered. Atthis point, the total volume of the filtrate was about 4.9 l. Thefiltrate was cooled to 5° C. and solid sodium carbonate was added toadjust the pH to 9.3.N-Carbethoxyphthalimide (263.04 g, 1.2 mol) wasadded in one portion and sodium carbonate was added as needed to keepthe pH at 9.3. After 2 hours at 5° C. followed by 3 hours at roomtemperature, the pH dropped to 8.5 and most of the reagents haddissolved. The reaction mixture was filtered, cooled to 5° C. andacidified to pH 2.3 with 6N hydrochloric acid. The precipitated solidwas collected by filtration and washed with 200 ml of cold water, thendried in vacuo to yield 220 g of title product.

EXAMPLE 28[4S-(4α,7α10aβ]-Octahydro-4-[[2-mercapto-3-(1-naphthalenyl)-1-oxopropyl]amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid,

a) (Acetylamino) (1-naphthalenylmethyl)propanedioic, diethyl ester

To a solution of sodium ethoxide (21% in ethanol, 4.613 gm, 67,8 mmol)in ethanol (100 ml) was added diethyl acetamidomalonate (14,74 gm, 67.8mmol), then 1-(bromomethyl)napthalene (10.0 gm, 45.2 mmol), The solutionwas stirred at room temperature for one hour. The reaction mixture wasthen concentrated to an orange oil. The oil was dissolved in ethylacetate and washed with 50% saturated ammonium chloride water and brine,then dried over sodium sulfate, filtered and concentrated to afford anorange solid. The solid was recrystallized from ethyl acetate and hexaneto afford beige crystals contaminated with diethylacetamido malonate.The solid was dissolved in 50% ethyl acetate in hexane and purified byflash chromatography on Merck silica gel in 50% ethyl acetate in hexane,Those fractions containing pure product were combined and concentratedto afford 10.225 g. of product as a white solid; m.p. 105°-108° C.;R_(f) =0.57 (50% ethyl acetate in hexane).

b) α-Amino-1-naphthalenepropanoic acid

A solution of the product from part (a) (16.182 gm., 47.5 mmol) wassuspended in 48% hydrogen bromide (100 ml) and refluxed under argon for14 hours. The hydrogen bromide salt of the product was filtered out ofsolution as a white solid, then taken up in hot (50° C.) water (500 ml)and the solution neutralized with concentrated ammonium hydroxide. Theproduct precipitated out of solution as a fine white solid. Uponfiltration and drying under high vacuum overnight (18 hours), 8.335 g.of product was obtained as a fluffy white solid; m.p. 264 ° C.

c) α-Bromo-1-naphthalenepropanoic acid

To a solution of the product from part (b) (4.000 g., 18.6 mmol) andpotassium bromide (7.63 g., 63.2 mmol) in 2.5N sulfuric acid (35 ml)kept at 0° C. was added sodium nitrite (1.92 g., 27.8 mmol) over onehour. The mixture was stirred for an additional hour at 0° C., then waswarmed to room temperature and stirred for 2.5 hours. The reactionmixture was then extracted with ether (3×). The ether layers werecombined and washed with water and brine, then dried over sodiumsulfate, filtered and concentrated to give an orange oil. The oil waspurified by flash chromatography on Merck silica gel in 70% ethylacetate in hexane with 1% acetic acid added to reduce tailing. Thosefractions containing the bromide were combined and concentrated toafford slightly contaminated product as an orange oil which solidifiedupon sitting overnight. R_(f) =0.40 (40% ethyl acetate in hexane with 1%acetic acid).

d) α-(Acetylthio)-1-naphthalenepropanoic acid

To a slurry of potassium thioacetate (0.912 g., 8.00 mmol) inacetonitrile (300 ml) at 0° C. was added the product from part (c)(2.030 g., 7.27 mmol) as a solution in acetonitrile (3 ml). The solutionwas stirred for one hour at 0° C., then was warmed to room temperatureand stirred for 15 hours. Potassium bromide was then filtered out of thereaction mixture and the filtrate concentrated to afford an orange oil.The oil was dissolved in ethyl acetate and washed with 10% potassiumbisulfate and brine, then dried over sodium sulfate, filtered andconcentrated to afford an orange oil. The oil was purified by flashchromatography on Merck silica gel in 50% ethyl acetate in hexane with1% acetic acid added to reduce tailing. Those fractions containingproduct were all contaminated with a compound with an R_(f) =0.43. Thosefractions were pooled and concentrated to give an orange oil. The crudeproduct was purified via the dicyclohexylamine salt by dissolving theorange oil in ether and adding an equivalent of dicyclohexylamine (1.32g., 7.27 mmol) to the solution. The dicyclohexylamine salt was obtainedin 2 crops of brown crystals (1.450 gm) still slightly contaminated withimpurity. The crystals were suspended in ethyl acetate and shaken with10% potassium bisulfate (3×). The organic layer was then washed withwater and brine, then dried over sodium sulfate filtered andconcentrated to afford 875 mg. of product as a yellow oil; R_(f) =0.40(40% ethyl acetate in hexane with 1% acetic acid).

e)[4S-(4α,7α10aβ)]-Octahydro-4-[[2-(acetylthio)-3-(1-naphthalenyl)-1-oxopropyl]amino]-5-oxo-7H-pyrido[2,1-b]-[1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of the racemic acid product from part (d) in methylenechloride and a solution of[4S-(4α,7α,10aβ)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester in methylene chloride are reacted in the presence oftriethylamine and benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate according to the procedure of Example 23(i) to givethe title product.

f)[4S-(4α,7α,10aβ)]-Octahydro-4-[[2-mercapto-3-(1-naphthalenyl)-1-oxopropyl]amino]-5-oxo-7H-pyrido-[2,1-b][1,3]-thiazepine-7-carboxylic acid

A solution of the product from part (e) in methanol is treated with 1Nsodium hydroxide according to the procedure of Example 23(j) to give thetitle product.

EXAMPLE 29[4S-[4α(R*),7α10aβ]]-Octahydro-4-[[2-(mercapto-1-oxo-3-(2-thienyl)propyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-[7-carboxylicacid,

a) (S)-α-(Acetylthio)-2-thiophenepropanoic acid

Potassium chloride (3.0 g., 40.1 mmol.) was added to a solution ofβ-(2-thienyl)-D-alanine (1.37 g., 8.03 mmol.) in 2.5N hydrochloric acid(25 ml.) at room temperature under argon. After stirring for 10 minutes,the resulting mixture was cooled to 0° C. and treated with sodiumnitrite (720 mg., 10.44 mmol.). After 2.5 hours, the reaction mixturewas warmed to room temperature and was stirred 1 hour. The mixture waspartitioned between water and ethyl acetate and the organic layer wasdried (sodium sulfate), filtered, and concentrated. The residue wasflash chromatographed (Merck silica gel) eluting with 1% acetic acid in3:1 hexane/ethyl acetate to give 760 mg. of(R)-α-chloro-2-thiophenecarboxylic acid as a yellow oil.

Cesium thioacetate (2.95 g., 14.19 mmol.) was added to a solutioncontaining the above chloride (750 mg., 4.73 mmol.) in dimethylformamide(15 ml.) at room temperature under argon. After stirring for 2 hours,the reaction mixture was partitioned between 10% potassium bisulfate andethyl acetate. The organic layer was washed with brine, dried (sodiumsulfate), filtered, and concentrated and the residue was flashchromatographed (Merck silica gel) eluting with 1% acetic acid in 4:1hexane/ethyl acetate to give 500 mg. of the title product as an oil. TLC(2% acetic acid in 3:1 ethyl acetate/hexane) Rf0.73.

[4S-[4α(R*),7α10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-(2-thienyl)propyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A solution of the acid product from part (a) in methylene chloride and asolution of[4S-(4α,7α,10aβ)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester in methylene chloride are reacted in the presence oftriethylamine and benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexoflurophosphate according to the procedure of Example 23(i) to givethe title product.

c)[4S-[4α(R*),7α10aβ]]-Octahydro-4-[[2-(mercapto-1-oxo-3-(2-thienyl)propyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid,

A solution of the product from part (b) in methanol is treated with 1Nsodium hydroxide according to the procedure of Example 23(j) to give thetitle product.

EXAMPLE 30[4S-[4α(S*),7α,10aβ]]-Octahydro-4[(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]-thiazepine-7-carboxylic acid

a) (R)-2-(Acetylthio)benzenepropanoic acid, dicylohexylamine salt

Following the procedure of Example 1(h) but substituting L-phenylalaninefor the D-phenylalanine, (R)-2-(acetylthio)benzenepropanoic acid,dicyclohexylamine salt was obtained.

b)[4S-[4α(S*),7α,10aβ]]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-phenylpropyl]amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

A stirred suspension of (R)-2-(acetylthio)benzenepropanoic acid,dicyclohexylamine salt (353.5 mg, 0.872 mmol) in ethyl acetate (5 ml)was washed with 5% potassium bisulfate solution (3×5 ml). The organicextracts were combined, washed with brine, dried (magnesium sulfate),filtered, concentrated, dried in vacuo and stripped twice from hexanesto obtain (R)-2-(acetylthio)benzenepropanoic acid as an oil.

The resulting free acid (181.4 mg, 0.809 mmol) was dissolved inmethylene chloride (2 ml) and stirred under nitrogen at 0° C. To thissolution was added a solution of[4S-(4α,7α10aβ)]-octahydro-4-amino-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester (200 mg, 0.774 mmol) in methylene chloride (6 ml),then triethylamine (0.113 ml, 0.813 mmol) and finallybenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(360 mg, 0.813 mmol). The reaction was stirred at 0° C. and then slowlyallowed to warm to room temperature. After a total of 20 hours, thereaction was concentrated in vacuo. The residue was dissolved in ethylacetate and the solution was washed with a 5% solution of potassiumbisulfate, a saturated solution of sodium bicarbonate, and brine. Theorganic layer was dried (magnesium sulfate), filtered, and concentratedto a yellow solid. Purification by flash chromatography (eluting with2:3 ethyl acetate/hexane) gave 261.7 mg of title product as a clear oil.

c)[4S-[4α(S*),7α,10aβ]]-Octahydro-4[-(2-mercapto-1-oxo-3-phenylpropyl)amino]-5-oxo-7H-pyrido[2,1-b][1,3]thiazepine-7-carboxylicacid

A solution of the product from part (b) (261.1 mg, 0.562 mmol) inmethanol (6 ml, deoxygenated via nitrogen bubbling) was cooled to 0° C.and treated with 1N sodium hydroxide (6 ml, deoxygenated via nitrogenbubbling). After stirring for one hour at 0° C. while purgingcontinuously with nitrogen, the reaction was warmed to room temperature.After stirring for 30 minutes at room temperature, a clear solution wasobtained. After 5.5 hours, the reaction was acidified, to pH 1 with 5%potassium bisulfate and extracted with ethyl acetate. The organic layerswere combined, washed with water and brine, dried (sodium sulfate),filtered and concentrated in vacuo. Purification by flash chromatography(6:0.01:3.99 ethyl acetate:acetic acid:hexane) gave 190 mg of titleproduct as a white solid; [α]_(D) =-87.5° (c=0.51, chloroform). TLC(6:0.01:3.99, ethyl acetate:acetic acid:hexane) R_(f) =0.20. HPLC: t_(R)=25.3 min; YMC S-3 ODS (C-18) 6.0×150 mm; 0% to 100% B:A, 30 min. lineargradient and 10 min. hold, 1.5 ml/min; A=90% water:methanol+0.2%phosphoric acid, B=90% methanol:water+0.2% phosphoric acid; 220 nm.

Anal. calc'd for C₁₉ H₂₄ O₄ N₂ S₂ •0.15 C₄ H₈ O₂ •0.15 C₇ H16•0.39 H₂ O:C, 55.89; H, 6.45; N, 6.31; S, 14.45 Found: C, 56.19; H, 6.50; N, 6.71;S, 13.96.

EXAMPLE 31[4S-(4α,7α,9aβ)]-Octahydro-4-[[2-mercapto-1-oxo-3-(4-thiazolyl)propyl]amino]-5-oxopyrrolo[2,1-b]-[1,3]-thiazepine-7-carboxylicacid

a) (R)-2-Amino-3-(4-thiazolyl)propanoic acid

A solution of 4N hydrochloric acid in dioxane (10 ml) was added to asolution of (R)-2-[[(1,1-dimethylethoxy)carbonyl]amino]-3-(4-thiazolyl)propanoic acid (2.0 g, 7.3 mmol) indioxane (2 ml). The reaction mixture was stirred at room temperature for3 hours, concentrated in vacuo and the residue was dissolved in water (3ml). The pH was adjusted to 6.5 with 1N sodium hydroxide and thissolution was passed through 20 ml of Dowex® AG50(H⁺). The column waseluted with water (250 ml) followed by 2% pyridine in water (300 ml).The product containing fractions were concentrated in vacuo to yield0.94 g of title product.

b) (R)-2-Bromo-3-(4-thiazolyl)propanoic acid

A solution of the product from part (a) (0.516 g, 3 mmol) and potassiumbromide (1.19 g, 10.1 mmol) in water (5.94 ml) and sulfuric acid (0.43ml) was stirred at -10° C. for 5 minutes followed by the portionwiseaddition of sodium nitrite (0.318 g, 4.61 mmol) over a 10 minute period.The reaction mixture was stirred an additional 10 minutes at 0° C. andat room temperature for one hour, and then extracted with ether (3×100ml). The ether extracts were washed with brine (2×20 ml), dried (sodiumsulfate), filtered, and concentrated in vacuo to yield 0.37 g of titleproduct; [α]_(D) =+37.35° (c=0.7, methanol). A second run was carriedout starting with 2.67 mmol of the product from part (a) using the sameprocedure to yield an additional 0.35 g of title product.

c) (S)-2-(Acetylthio)-3-(4-thiazolyl)propanoic acid

The product from part (b) (0.72 g, 3.05 mmol) and potassium thioacetate(0.35 g, 3.05 mmol) were stirred in acetonitrile (9 ml) overnight atroom temperature and at 30° C. for one hour. The reaction mixture wasdiluted with ethyl acetate (100 ml) and filtered. The filtrate wasconcentrated in vacuo. The residue was redissolved in ethyl acetate (100ml), washed with water (2×50 ml) and brine (20 ml), dried (sodiumsulfate), filtered and concentrated in vacuo to yield 0.52 g of titleproduct; [α]_(D) =-15.89° (c=0.6, methanol).

d)[4S-(4α,7α,9aβ)]-Octahydro-4-[[2-(acetylthio)-1-oxo-3-(4-thiazolyl)propyl]amino]-5-oxopyrrolo-[2,1-b][1,3]thiazepine-7-carboxylicacid, methyl ester

[4S-(4α,7α,9aβ)]-4-Amino-octahydro-5-oxo-pyrrolo[2,1-b][1,3]thiazepine-7-carboxylicacid methyl ester, p-toluenesulfonic acid salt [0.3 67 g, 0.882 mmol,prepared from the material described in Example 5(d)] was dissolved inmethylene chloride (5 ml) 0° C., followed by the addition oftriethylamine (0.12 ml, 0.868 mmol). The product from part (c) (0.2 g,0.865 mmol) was added to this solution followed by a second portion oftriethylamine (0.12 ml, 0. 865 mmol).Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(0,383 g, 0,865 mmol) was added and the reaction mixture was stirred at0° C. for one hour and a room temperature for 4 hours. The reactionmixture was concentrated in vacuo and the residue was dissolved in ethylacetate (60 ml). The organic extract was washed with 5% aqueouspotassium bisulfate (10 ml) and brine (2×10 ml), dried (sodium sulfate),filtered and concentrated in vacuo. This crude material waschromatographed through 100 g of Merck silica gel using 0.2% methanol inethyl acetate. The fractions enriched in the slower isomer wereconcentrated in vacuo to yield 0.134 g of title product.

e)[4S-(4α,7α,9aβ)]-Octahydro-4-[[2-mercapto-1-oxo-3-(4-thiazolyl)propyl]amino]-5-oxopyrrolo-[2,1-b][1,3]thiazepine-7-carboxylicacid

The product from part (d) (0.135 g, 0.29 mmol) was dissolved in methanol(3 ml) and argon was bubbled into the solution for 30 minutes at 0° C.1N Sodium hydroxide (1.32 ml) also purged with argon was added to theabove solution and the reaction mixture was stirred at 0° C. with argonbubbling through the solution for one hour and at room temperature for 2hours. The reaction was quenched by the addition of 5% aqueous potassiumbisulfate (20 ml) and the organics were extracted with ethyl acetate(3×50 ml). The ethyl acetate solution was washed with brine, dried(sodium sulfate), filtered, and concentrated in vacuo. The concentratewas chromatographed through 40 g of Merck silica gel using chloroformcontaining 5% methanol and 0.5% acetic acid. The appropriate fractionswere combined, concentrated and partitioned between 20 ml of ethylacetate and 5% aqueous potassium bisulfate. The ethyl acetate solutionwas washed with water and brine, dried (sodium sulfate), andconcentrated in vacuo. The residue was lyophilized from dioxane (4 ml)to yield 36 mg of title product as a 70:30 mixture of isomers; m.p.95°-115° C.; [α]_(D) =-191.7° (c=0.06, chloroform). TLC(chloroform:methanol:acetic acid, 8:2:0.2) R_(f) =0.59. HPLC: t_(R)=3.06 min; YMC S-3 ODS (C-18) 6.0×150 mm, 3μ end capped column,isocratic 60% aqueous methanol containing 0.2% phosphoric acid, 25 min,1.5 ml/min. (95.4%).

Anal. calc'd for C₁₅ H₁₉ N₃ O₄ S₃ •0.2 C₄ H₈ O₂ •0.9 H₂ O: C, 43.59; H,5.19; N, 9.65; S, 22.09 Found: C, 43.54; H, 4,89; N, 9.44; S, 21.90.

EXAMPLE 32

1000 tablets each containing the following ingredients:

    ______________________________________                                        [4S-[4α(R*),7α,10aβ]]-Octahydro-                                                      100    mg.                                           4-[(2-mercapto-1-oxo-3-phenylpropyl)-                                         amino]-5-oxo-7H-pyrido[2,1-b][1,3]thia-                                       zepine-7-carboxylic acid                                                      Cornstarch               100    mg.                                           Gelatin                  20     mg.                                           Avicel(microcrystalline cellulose)                                                                     50     mg.                                           Magnesium stearate       5      mg.                                                                    275    mg.                                           ______________________________________                                    

are prepared from sufficient bulk quantities by mixing the product ofExample 3 and cornstarch with an aqueous solution of the gelatin. Themixture is dried and ground to a fine powder. The Avicel and then themagnesium stearate are admixed with granulation. The mixture is thencompressed in a tablet press to form 1000 tablets each containing 100mg. of active ingredient.

In a similar manner, tablets containing 200 mg. of the product ofExamples 1 2, 4 to 23, 25, and 28 to 31 can be prepared.

Similar procedures can be employed to form tablets or capsulescontaining from 10 mg. to 500 mg. of active ingredient.

What is claimed is:
 1. A process for preparing a compound of the formula##STR85## wherein P₁ is a nitrogen protecting group and R₆ is alkyl,substituted alkyl, or benzyl which comprises:a) reacting L-methionine tointroduce the P₁ protecting group; b) esterifying the product from part(a) by treating with an alcohol, alkyl-OH, in the presence of an acidcatalyst to give ##STR86## c) oxidizing the ester product of part (b) togive ##STR87## d) treating the sulfoxide product of part (c) with anacid anhydride of the formula ##STR88## to give ##STR89## e) treatingthe product of part (d) with an alkali metal hydroxide followed byremoval of formaldehyde, and treatment with an acid anhydride as definedabove or an acid halide halo-C(O)--R₆ to give the desired product.
 2. Aprocess of claim 1 wherein the oxidizing reaction in step (c) isperformed by using N-chlorosuccinimide.
 3. A process of claim 1 whereinP₁ is ##STR90## and R₆ is methyl which comprises: a) reactingL-methionine with benzylchloroformate to give ##STR91## b) treating theproduct from part (a) with methanol in the presence of p-toluenesulfonicacid monohydrate to give ##STR92## c) oxidizing the product of part (b)by treating with N-chlorosuccinimide in aqueous methanol to give##STR93## d) treating the sulfoxide product of part (c) with aceticanhydride to give ##STR94## e) treating the product of part (d) withpotassium hydroxide followed by removal of formaldehyde, and treatmentwith acetic anhydride to give the desired product.